BEGIN:VCALENDAR VERSION:2.0 PRODID:-//CERN//INDICO//EN BEGIN:VEVENT SUMMARY:Fragment Based Active Site Exploration of Polyurethane Degrading E nzymes for Structure-guided Protein Engineering DTSTART;VALUE=DATE-TIME:20250922T174000Z DTEND;VALUE=DATE-TIME:20250922T175000Z DTSTAMP;VALUE=DATE-TIME:20260524T212503Z UID:indico-contribution-583-1866@lindico453.srv.lu.se DESCRIPTION:Speakers: Deniz Bicer (Aarhus University)\nPolyurethane (PU) p lastics are extermyl durable and hard to recycle by convential methods due to\ncrosslinked and branched molecular structure. Recent discoveries of e nzyme that arget carbamate\n(urethane) bond in the PU provide alternative way of PU recycling by means of biocatalytic degradation.\nEnZync center h as been established to discover\, characterize and engineer novel enzymes for PU\ndegradation. So far we have discovered several enzymes by computat ional searches and experimental\nways. The critical steps to establish rat ional basis of protein engineering campaing is the understanding\nof Plast ic-enzyme interactions at molecular level. To pursue structural characteri zation of “PURases”\nwe have two different but complemantry methods to characteirze active site of PURases: i) Fragment\nBased Active site Explo ration (FASE) of PURases by using small molecule fragment libraries and\ns oluble PU analogs and ii) time-resolved serial crystallography by cryocapt uring catalytic intermideate\n(for slow-milisecond kinetics) and ambient t emperature (fast-microsecond) crystallography. We have\ncompleted FASE app roaches for one of our patented enzymes and now we are strating to perform serial\ncrystallography approach with promising preliminary data.\n\nAuth ors: Deniz Bicer\, Laura Rotilio\, Daniel Otzen & Jens Preben Morth\n\nhtt ps://lindico453.srv.lu.se/event/583/contributions/1866/ LOCATION:LINXS at The Loop URL:https://lindico453.srv.lu.se/event/583/contributions/1866/ END:VEVENT BEGIN:VEVENT SUMMARY:MicroMAX - A beamline with time-resolved macromolecular crystallog raphy capabilities at the MAX IV Laboratory DTSTART;VALUE=DATE-TIME:20250922T173000Z DTEND;VALUE=DATE-TIME:20250922T174000Z DTSTAMP;VALUE=DATE-TIME:20260524T212503Z UID:indico-contribution-583-1865@lindico453.srv.lu.se DESCRIPTION:Speakers: Jie Nan et al (MAX IV Laboratory)\nThe rise of 4th g eneration sources\, including the MAX IV Laboratory 3 GeV ring\, has enabl ed new possibilities to study dynamics using crystallography. The MicroMAX beamline is a new beamline focussed on providing optimal X-ray characteri stics for serial (SSX) and time-resolved (TR-SSX) crystallography at MAX I V [1]. The beamline emphasizes a flexible sample environment for standard and bespoke experimental setups while also supporting high-throughput sing le crystal data collections at the BioMAX beamline which has operated sinc e 2017 [2]. \nThe MicroMAX user program opened in May 2024 and has perform ed experiments with SPINE-based fixed targets\, high-viscosity extrusion a nd microfluidics and single-crystal oscillation data collections. Sample h andling and positioning is supported by the MD3-up micro diffractometer\, Oxford cryojet\, and ISARA automated sample mounting platform (including c rystallization plates). Time resolved techniques are enabled by a nanoseco nd pump laser (210-2600 nm)\, Celerotron X-ray chopper (0\,8-70% duty cycl e) and one of either an Eiger2 X 9M CdTe photon counting hybrid pixel dete ctor or Jungfrau 9M Si integrating hybrid pixel detector (on-loan from PSI ). \nOptical elements allow for a beamline flux from 10^13 photons/s (0.1% bandwidth double crystal monochromator) to more than 10^14 photons/s (1% bandwidth multilayer monochromator) with an optimal 1x1 μm beam focus usi ng beryllium lenses/K-B mirrors. Beamline controls are from within MXCuBE\ , with additional live feedback and CrystFEL autoprocessing pipelines to p rovide immediate feedback and rapid map generation. Sample pre-characteriz ation is supported by an offline laser and spectroscopy lab in the seconda ry experimental hutch and dedicated sample environment and preparation lab s.\nHere we present the current status of MicroMAX beamline and recent dev elopments in sample preparation and data handling under a variety of exper imental contexts. This work emphasizes the technical developments for a hi ghly flexible TR-SSX end station in context of SSX/TR-SSX experiments alre ady being conducted by the MicroMAX user community.\n\nMicroMAX is funded by the Novo Nordisk Foundation under the grant number NNF17CC0030666.\n\n[ 1] Gonzalez\, A.\, Krojer\, T.\, Nan\, J.\, Bjelcic\, M.\, Aggarwal\, S.\, Gorgisyan\, I.\, Milas\, M.\, Eguiraun\, M.\, Casadei\, C.\, Chenchiliyan \, M.\, Jurgilaitis\, A.\, Kroon\, D.\, Ahn\, B.\, Ekstrom\, J. C.\, Aurel ius\, O.\, Lang\, D.\, Ursby\, T. & Thunnissen\, M. M. G. M. (2025). J. Sy nchrotron Rad. 32.\n[2] Shilova\, A.\, Lebrette\, H.\, Aurelius\, O.\, Nan \, J.\, Welin\, M.\, Kovacic\, R.\, Ghosh\, S.\, Safari\, C.\, Friel\, R. J.\, Milas\, M.\, Matej\, Z.\, Högbom\, M.\, Brändén\, G.\, Kloos\, M.\ , Shoeman\, R. L.\, Doak\, B.\, Ursby\, T.\, Håkansson\, M.\, Logan\, D. T. & Mueller U. (2020). J. Synchrotron Rad.\, 27\, 1095.\n\nhttps://lindic o453.srv.lu.se/event/583/contributions/1865/ LOCATION:LINXS at The Loop URL:https://lindico453.srv.lu.se/event/583/contributions/1865/ END:VEVENT BEGIN:VEVENT SUMMARY:Laser and Spectroscopic Capabilities at MicroMAX DTSTART;VALUE=DATE-TIME:20250922T172000Z DTEND;VALUE=DATE-TIME:20250922T173000Z DTSTAMP;VALUE=DATE-TIME:20260524T212503Z UID:indico-contribution-583-1864@lindico453.srv.lu.se DESCRIPTION:Speakers: Manoop Chenchiliyan (MAX IV Laboratory)\, Sofia M. K apetanaki (MAX IV Laboratory)\nSpectroscopic techniques provide a powerful means of obtaining detailed information on the structural and dynamic pro perties of proteins in solution and in crystallo (proteins and enzymes are active in the crystalline state). Structural data obtained by X-ray cryst allography is strengthened by insights from complementary approaches like UV/vis\, fluorescence and (resonance) Raman measurements.\n\nhttps://lindi co453.srv.lu.se/event/583/contributions/1864/ LOCATION:LINXS at The Loop URL:https://lindico453.srv.lu.se/event/583/contributions/1864/ END:VEVENT BEGIN:VEVENT SUMMARY:Serial femtosecond crystallography of high-valent metal sites and protein radicals DTSTART;VALUE=DATE-TIME:20250923T084500Z DTEND;VALUE=DATE-TIME:20250923T091500Z DTSTAMP;VALUE=DATE-TIME:20260524T212503Z UID:indico-contribution-583-1848@lindico453.srv.lu.se DESCRIPTION:Speakers: Martin Högbom (Stockholm University)\nHögbom\, M 1 \n\n1. Department of Biochemistry and Biophysics\, Stockholm University\, 10691 Stockholm\, Sweden.\n\nHigh resolution structure determination metho ds suffer from problems with radiation damage. This is particularly proble matic for radiation sensitive states such as high-valent metal sites and r adicals. From a chemical perspective this means that some of the most rele vant states for catalysis in many enzyme systems are inaccessible to stand ard structure determination regimes. \nIn close collaboration with scienti sts at the LCLS and the LBNL we utilize a conveyor-belt sample injector th at allows micrometer-sized crystals to be manipulated in various ways\, in cluding oxygen incubation for a defined period of time\, before exposure t o the free-electron laser X-ray beam [1]. This setup allows varying the ti me for intermediate trapping while the use of femtosecond XFEL crystallogr aphy eliminates the effect of X-ray photoreduction on obtained data. Simul taneous XES also allows in situ oxidation state determination of probed in termediates for metalloprotein systems. \nThis setup and its use to obtain high-resolution global geometric structures of high-valent intermediates will be discussed\, as well as our recent progress defining radiation unda maged structures of methane monooxygenase [2] and ribonucleotide reductase R2 proteins [3\,4] including the catalytic radical state [5].\n\n\nAcknow ledgments: This work was funded by the Knut and Alice Wallenberg Foundatio n\, the Swedish Research Council and the European Research Council (ERC).\ n\nReferences\n[1] F.D. Fuller et al. Nature Methods\, 14(4):443-449 (2017 )\n[2] Srinivas V. et al. J Am Chem Soc\, 142:14249-14266 (2020)\n[3] Srin ivas V. et al. Nature\, 563:416-420 (2018)\n[4] John J. et al. Elife\, 11: e79226 (2022)\n[5] Lebrette H. et al. Science\, 382:109-113 (2023)\n\nhttp s://lindico453.srv.lu.se/event/583/contributions/1848/ LOCATION:LINXS at The Loop URL:https://lindico453.srv.lu.se/event/583/contributions/1848/ END:VEVENT BEGIN:VEVENT SUMMARY:Data analysis at MicroMAX DTSTART;VALUE=DATE-TIME:20250923T121000Z DTEND;VALUE=DATE-TIME:20250923T123000Z DTSTAMP;VALUE=DATE-TIME:20260524T212503Z UID:indico-contribution-583-1853@lindico453.srv.lu.se DESCRIPTION:Speakers: Cecilia Casadei (MAX IV Laboratory)\nThe rise of 4th generation sources\, including the MAX IV Laboratory 3 GeV ring\, has ena bled new possibilities to study dynamics using crystallography. The MicroM AX beamline is a new beamline focussed on providing optimal X-ray characte ristics for serial (SSX) and time-resolved (TR-SSX) crystallography at MAX IV [1]. The beamline emphasizes a flexible sample environment for standar d and bespoke experimental setups while also supporting high-throughput si ngle crystal data collections at the BioMAX beamline which has operated si nce 2017 [2]. \nThe MicroMAX user program opened in May 2024 and has perfo rmed experiments with SPINE-based fixed targets\, high-viscosity extrusion and microfluidics and single-crystal oscillation data collections. Sample handling and positioning is supported by the MD3-up micro diffractometer\ , Oxford cryojet\, and ISARA automated sample mounting platform (including crystallization plates). Time resolved techniques are enabled by a nanose cond pump laser (210-2600 nm)\, Celerotron X-ray chopper (0\,8-70% duty cy cle) and one of either an Eiger2 X 9M CdTe photon counting hybrid pixel de tector or Jungfrau 9M Si integrating hybrid pixel detector (on-loan from P SI). \nOptical elements allow for a beamline flux from 10^13 photons/s (0. 1% bandwidth double crystal monochromator) to more than 10^14 photons/s (1 % bandwidth multilayer monochromator) with an optimal 1x1 μm beam focus u sing beryllium lenses/K-B mirrors. Beamline controls are from within MXCuB E\, with additional live feedback and CrystFEL autoprocessing pipelines to provide immediate feedback and rapid map generation. Sample pre-character ization is supported by an offline laser and spectroscopy lab in the secon dary experimental hutch and dedicated sample environment and preparation l abs.\nHere we present the current status of MicroMAX beamline and recent d evelopments in sample preparation and data handling under a variety of exp erimental contexts. This work emphasizes the technical developments for a highly flexible TR-SSX end station in context of SSX/TR-SSX experiments al ready being conducted by the MicroMAX user community.\n\nMicroMAX is funde d by the Novo Nordisk Foundation under the grant number NNF17CC0030666.\n\ n[1] Gonzalez\, A.\, Krojer\, T.\, Nan\, J.\, Bjelcic\, M.\, Aggarwal\, S. \, Gorgisyan\, I.\, Milas\, M.\, Eguiraun\, M.\, Casadei\, C.\, Chenchiliy an\, M.\, Jurgilaitis\, A.\, Kroon\, D.\, Ahn\, B.\, Ekstrom\, J. C.\, Aur elius\, O.\, Lang\, D.\, Ursby\, T. & Thunnissen\, M. M. G. M. (2025). J. Synchrotron Rad. 32.\n[2] Shilova\, A.\, Lebrette\, H.\, Aurelius\, O.\, N an\, J.\, Welin\, M.\, Kovacic\, R.\, Ghosh\, S.\, Safari\, C.\, Friel\, R . J.\, Milas\, M.\, Matej\, Z.\, Högbom\, M.\, Brändén\, G.\, Kloos\, M .\, Shoeman\, R. L.\, Doak\, B.\, Ursby\, T.\, Håkansson\, M.\, Logan\, D . T. & Mueller U. (2020). J. Synchrotron Rad.\, 27\, 1095.\n\nhttps://lind ico453.srv.lu.se/event/583/contributions/1853/ LOCATION:LINXS at The Loop URL:https://lindico453.srv.lu.se/event/583/contributions/1853/ END:VEVENT BEGIN:VEVENT SUMMARY:Sample delivery at MicroMAX DTSTART;VALUE=DATE-TIME:20250923T115000Z DTEND;VALUE=DATE-TIME:20250923T121000Z DTSTAMP;VALUE=DATE-TIME:20260524T212503Z UID:indico-contribution-583-1852@lindico453.srv.lu.se DESCRIPTION:Speakers: Dean Lang (MAX IV Laboratory)\nThe rise of 4th gener ation sources\, including the MAX IV Laboratory 3 GeV ring\, has enabled n ew possibilities to study dynamics using crystallography. The MicroMAX bea mline is a new beamline focussed on providing optimal X-ray characteristic s for serial (SSX) and time-resolved (TR-SSX) crystallography at MAX IV [1 ]. The beamline emphasizes a flexible sample environment for standard and bespoke experimental setups while also supporting high-throughput single c rystal data collections at the BioMAX beamline which has operated since 20 17 [2]. \nThe MicroMAX user program opened in May 2024 and has performed e xperiments with SPINE-based fixed targets\, high-viscosity extrusion and m icrofluidics and single-crystal oscillation data collections. Sample handl ing and positioning is supported by the MD3-up micro diffractometer\, Oxfo rd cryojet\, and ISARA automated sample mounting platform (including cryst allization plates). Time resolved techniques are enabled by a nanosecond p ump laser (210-2600 nm)\, Celerotron X-ray chopper (0\,8-70% duty cycle) a nd one of either an Eiger2 X 9M CdTe photon counting hybrid pixel detector or Jungfrau 9M Si integrating hybrid pixel detector (on-loan from PSI). \ nOptical elements allow for a beamline flux from 10^13 photons/s (0.1% ban dwidth double crystal monochromator) to more than 10^14 photons/s (1% band width multilayer monochromator) with an optimal 1x1 μm beam focus using b eryllium lenses/K-B mirrors. Beamline controls are from within MXCuBE\, wi th additional live feedback and CrystFEL autoprocessing pipelines to provi de immediate feedback and rapid map generation. Sample pre-characterizatio n is supported by an offline laser and spectroscopy lab in the secondary e xperimental hutch and dedicated sample environment and preparation labs.\n Here we present the current status of MicroMAX beamline and recent develop ments in sample preparation and data handling under a variety of experimen tal contexts. This work emphasizes the technical developments for a highly flexible TR-SSX end station in context of SSX/TR-SSX experiments already being conducted by the MicroMAX user community.\n\nMicroMAX is funded by t he Novo Nordisk Foundation under the grant number NNF17CC0030666.\n\n[1] G onzalez\, A.\, Krojer\, T.\, Nan\, J.\, Bjelcic\, M.\, Aggarwal\, S.\, Gor gisyan\, I.\, Milas\, M.\, Eguiraun\, M.\, Casadei\, C.\, Chenchiliyan\, M .\, Jurgilaitis\, A.\, Kroon\, D.\, Ahn\, B.\, Ekstrom\, J. C.\, Aurelius\ , O.\, Lang\, D.\, Ursby\, T. & Thunnissen\, M. M. G. M. (2025). J. Synchr otron Rad. 32.\n[2] Shilova\, A.\, Lebrette\, H.\, Aurelius\, O.\, Nan\, J .\, Welin\, M.\, Kovacic\, R.\, Ghosh\, S.\, Safari\, C.\, Friel\, R. J.\, Milas\, M.\, Matej\, Z.\, Högbom\, M.\, Brändén\, G.\, Kloos\, M.\, Sh oeman\, R. L.\, Doak\, B.\, Ursby\, T.\, Håkansson\, M.\, Logan\, D. T. & Mueller U. (2020). J. Synchrotron Rad.\, 27\, 1095.\n\nhttps://lindico453 .srv.lu.se/event/583/contributions/1852/ LOCATION:LINXS at The Loop URL:https://lindico453.srv.lu.se/event/583/contributions/1852/ END:VEVENT BEGIN:VEVENT SUMMARY:MicroMAX - Overview DTSTART;VALUE=DATE-TIME:20250923T113000Z DTEND;VALUE=DATE-TIME:20250923T115000Z DTSTAMP;VALUE=DATE-TIME:20260524T212503Z UID:indico-contribution-583-1851@lindico453.srv.lu.se DESCRIPTION:Speakers: Jie Nan (MAX IV Laboratory)\nThe rise of 4th generat ion sources\, including the MAX IV Laboratory 3 GeV ring\, has enabled new possibilities to study dynamics using crystallography. The MicroMAX beaml ine is a new beamline focussed on providing optimal X-ray characteristics for serial (SSX) and time-resolved (TR-SSX) crystallography at MAX IV [1]. The beamline emphasizes a flexible sample environment for standard and be spoke experimental setups while also supporting high-throughput single cry stal data collections at the BioMAX beamline which has operated since 2017 [2]. \nThe MicroMAX user program opened in May 2024 and has performed exp eriments with SPINE-based fixed targets\, high-viscosity extrusion and mic rofluidics and single-crystal oscillation data collections. Sample handlin g and positioning is supported by the MD3-up micro diffractometer\, Oxford cryojet\, and ISARA automated sample mounting platform (including crystal lization plates). Time resolved techniques are enabled by a nanosecond pum p laser (210-2600 nm)\, Celerotron X-ray chopper (0\,8-70% duty cycle) and one of either an Eiger2 X 9M CdTe photon counting hybrid pixel detector o r Jungfrau 9M Si integrating hybrid pixel detector (on-loan from PSI). \nO ptical elements allow for a beamline flux from 10^13 photons/s (0.1% bandw idth double crystal monochromator) to more than 10^14 photons/s (1% bandwi dth multilayer monochromator) with an optimal 1x1 μm beam focus using ber yllium lenses/K-B mirrors. Beamline controls are from within MXCuBE\, with additional live feedback and CrystFEL autoprocessing pipelines to provide immediate feedback and rapid map generation. Sample pre-characterization is supported by an offline laser and spectroscopy lab in the secondary exp erimental hutch and dedicated sample environment and preparation labs.\nHe re we present the current status of MicroMAX beamline and recent developme nts in sample preparation and data handling under a variety of experimenta l contexts. This work emphasizes the technical developments for a highly f lexible TR-SSX end station in context of SSX/TR-SSX experiments already be ing conducted by the MicroMAX user community.\n\nMicroMAX is funded by the Novo Nordisk Foundation under the grant number NNF17CC0030666.\n\n[1] Gon zalez\, A.\, Krojer\, T.\, Nan\, J.\, Bjelcic\, M.\, Aggarwal\, S.\, Gorgi syan\, I.\, Milas\, M.\, Eguiraun\, M.\, Casadei\, C.\, Chenchiliyan\, M.\ , Jurgilaitis\, A.\, Kroon\, D.\, Ahn\, B.\, Ekstrom\, J. C.\, Aurelius\, O.\, Lang\, D.\, Ursby\, T. & Thunnissen\, M. M. G. M. (2025). J. Synchrot ron Rad. 32.\n[2] Shilova\, A.\, Lebrette\, H.\, Aurelius\, O.\, Nan\, J.\ , Welin\, M.\, Kovacic\, R.\, Ghosh\, S.\, Safari\, C.\, Friel\, R. J.\, M ilas\, M.\, Matej\, Z.\, Högbom\, M.\, Brändén\, G.\, Kloos\, M.\, Shoe man\, R. L.\, Doak\, B.\, Ursby\, T.\, Håkansson\, M.\, Logan\, D. T. & M ueller U. (2020). J. Synchrotron Rad.\, 27\, 1095.\n\nhttps://lindico453.s rv.lu.se/event/583/contributions/1851/ LOCATION:LINXS at The Loop URL:https://lindico453.srv.lu.se/event/583/contributions/1851/ END:VEVENT BEGIN:VEVENT SUMMARY:Time-resolved X-ray crystallography on membrane proteins: Watching ions moving in time and space DTSTART;VALUE=DATE-TIME:20250923T070000Z DTEND;VALUE=DATE-TIME:20250923T073000Z DTSTAMP;VALUE=DATE-TIME:20260524T212503Z UID:indico-contribution-583-1845@lindico453.srv.lu.se DESCRIPTION:Speakers: Przemyslaw Nogly (Jagiellonian University in Krakow) \nP. Nogly\nDioscuri Centre for Structural Dynamics of Receptors\, Faculty of Biochemistry\, Biophysics and Biotechnology\, Jagiellonian University\ , Kraków\, Poland\; przemyslaw.nogly@uj.edu.pl \n\nChloride transport is an essential process maintaining ion balance across cell membranes\, cell growth\, and neuronal action potentials. However\, the molecular mechanism of the transport remains elusive. Among chloride transporters\, light-dri ven rhodopsins have gained attention as optogenetic tools to manipulate ne uronal signaling. We combined time-resolved serial crystallography at XFEL and synchrotron to provide a comprehensive view of chloride-pumping rhodo psin's structural dynamics and molecular mechanism throughout the transpor t cycle from 10 ps to 50 ms [1]. We traced transient anion binding sites\, obtained evidence for the mechanism of light energy utilization in transp ort\, and identified steric and electrostatic molecular gates ensuring uni directional transport. These structural insights provided the basis for mu tagenesis and functional study of the mechanistic features enabling finely controlled chloride transport across the cell membrane.\nFurthermore\, ou r recent study of a distinct photoreceptor\, Light-Oxygen-Voltage (LOV) do main\, will be introduced. The first insights into the structural dynamics of LOV photoactivation will be presented\, providing the basis for propos ing a molecular mechanism of a covalent thioether bond formation between a flavin mononucleotide cofactor and a reactive cysteine\, Cys57 (unpublish ed).\n\n[1] Mous\, S. et al. Science 375 (2022) 845\n\nhttps://lindico453. srv.lu.se/event/583/contributions/1845/ LOCATION:LINXS at The Loop URL:https://lindico453.srv.lu.se/event/583/contributions/1845/ END:VEVENT BEGIN:VEVENT SUMMARY:Posters & mingle food (Poster list with abstracts below) DTSTART;VALUE=DATE-TIME:20250922T151500Z DTEND;VALUE=DATE-TIME:20250922T153000Z DTSTAMP;VALUE=DATE-TIME:20260524T212503Z UID:indico-contribution-583-1834@lindico453.srv.lu.se DESCRIPTION:https://lindico453.srv.lu.se/event/583/contributions/1834/ LOCATION:LINXS at The Loop URL:https://lindico453.srv.lu.se/event/583/contributions/1834/ END:VEVENT BEGIN:VEVENT SUMMARY:Electrostatically Gated Enzyme Dynamics During Catalysis DTSTART;VALUE=DATE-TIME:20250922T143000Z DTEND;VALUE=DATE-TIME:20250922T150000Z DTSTAMP;VALUE=DATE-TIME:20260524T212503Z UID:indico-contribution-583-1831@lindico453.srv.lu.se DESCRIPTION:Speakers: Mark Wilson (University of Nebraska–Lincoln)\nAbst ract: Enzyme catalysis is essential for life and is a central phenomenon i n biochemistry. The advent of time-resolved serial crystallography\, initi ally enabled by X-ray free electron lasers (XFELs) and now expanding to sy nchrotron X-ray sources\, allows enzyme catalysis to be observed catalysis in real time\, in near-physiological conditions\, and at atomic resolutio n. I will describe our work using mix-and-inject serial crystallography (M ISC) to observe catalysis by isocyanide hydratase (ICH). MISC allowed us to observe formation of an unusual thioimidate intermediate and to watch I CH’s conformational dynamics respond to changes in active site ionizatio n during catalysis. We also used an engineered ICH mutant to enrich for r are conformations during catalysis\, permitting a clearer view of later st eps in the reaction. ICH exemplifies a class of enzymes whose non-equilib rium dynamics are gated by changes in active site electrostatics\, which i s a potentially common enzymological phenomenon.\n\nhttps://lindico453.srv .lu.se/event/583/contributions/1831/ LOCATION:LINXS at The Loop URL:https://lindico453.srv.lu.se/event/583/contributions/1831/ END:VEVENT BEGIN:VEVENT SUMMARY:Time-resolved crystallography of light-driven ion transporters at synchrotrons and free electron lasers DTSTART;VALUE=DATE-TIME:20250922T132500Z DTEND;VALUE=DATE-TIME:20250922T135500Z DTSTAMP;VALUE=DATE-TIME:20260524T212503Z UID:indico-contribution-583-1830@lindico453.srv.lu.se DESCRIPTION:Speakers: Kirill Kovalev (EMBL-Hamburg)\nMicrobial rhodopsins constitute a large superfamily of light-sensitive membrane proteins\, whic h are vital for numerous microorganisms on Earth\, and for 20 years also b eing most-actively used in neuroscience and medicine in the biotechnology named optogenetics. The major role for optogenetics is played by ion-trans porting rhodopsins. Understanding of their molecular mechanisms of functio ning can not only contribute to the fundamental biological knowledge of th eir roles in native host\, but can enable their optimization towards routi ne use in medical application\, such as restoration of eye vision and hear ing. I will report on our recent advances in investigations of light-drive n ion pumps\, with particular focus on applying time-resolved serial cryst allography at X-ray free electron lasers and synchrotrons to obtain molecu lar movies of the proteins in action. These include tracking of ultrafast changes and slower but more prominent rearrangements in a new type of sodi um transporters\, as well as in inward proton pumps. I will also discuss t he optimization of sample delivery for more efficient time-resolved crysta llography experiments and its implication for other systems\, such as near -infrared-absorbing microbial rhodopsins.\n\nhttps://lindico453.srv.lu.se/ event/583/contributions/1830/ LOCATION:LINXS at The Loop URL:https://lindico453.srv.lu.se/event/583/contributions/1830/ END:VEVENT BEGIN:VEVENT SUMMARY:From Static to Dynamic: Evolving Methods in Protein Structure Dete rmination DTSTART;VALUE=DATE-TIME:20250922T124000Z DTEND;VALUE=DATE-TIME:20250922T131000Z DTSTAMP;VALUE=DATE-TIME:20260524T212503Z UID:indico-contribution-583-1828@lindico453.srv.lu.se DESCRIPTION:Speakers: Aina Cohen (Stanford Synchrotron Radiation Lightsour ce)\n(representing the entire SSRL-SMB team)\nStructural Molecular Biology (SMB)\, Stanford Synchrotron Radiation Lightsource (SSRL)\, SLAC National Accelerator Laboratory\, Stanford University\, Menlo Park\, United States of America\n\nStructural biologists are undertaking increasingly challeng ing projects including the study of membrane proteins and complex multi-co mponent machines. Structural investigations are also transitioning beyond solving a single static structure\, to the application of a series of sequ ential structural snapshots to provide details of the atomic positions and motions that define the relationships involved in molecular recognition\, transition state stabilization\, and other aspects of the biocatalytic pr ocess. The success of these experiments requires careful optimization of s amples and experimental setups\, often involving multiple experiments at t he laboratory bench and the beamline\, where automation serves as an enabl ing technology to efficiently deliver multiple crystals and meet stringent timing requirements. \nDevelopments at SSRL and LCLS-MFX will be presente d that tackle challenges involved in the study of metalloenzymes\, the use of small and radiation-sensitive crystals\, and to perform time-resolved crystallography. To facilitate the handling and optimization of delicate c rystals\, new in situ crystallization and remote data collection schemes h ave been released that avoid direct manipulation of crystals\, support rob otic sample exchange\, and allow full rotational access of the sample in a controlled humidity environment. By simplifying crystal handling and tran sport at near-physiological temperatures\, these technologies remove barri ers to enable more widespread use of serial crystallography methods for st udies of metalloenzyme structure and protein dynamics. Strategies for time -resolved measurements and data analysis tools that provide rapid feedback for experimental optimization during fast-paced experiments will also be described.\n\nhttps://lindico453.srv.lu.se/event/583/contributions/1828/ LOCATION:LINXS at The Loop URL:https://lindico453.srv.lu.se/event/583/contributions/1828/ END:VEVENT BEGIN:VEVENT SUMMARY:The potential for serial crystallography in drug discovery DTSTART;VALUE=DATE-TIME:20250922T114000Z DTEND;VALUE=DATE-TIME:20250922T121000Z DTSTAMP;VALUE=DATE-TIME:20260524T212503Z UID:indico-contribution-583-1825@lindico453.srv.lu.se DESCRIPTION:Speakers: Helena Käck ()\nAndreas Dunge (1\,2)\, Gabrielle We hlander (2)\, Gisela Brändén(2) and Helena Käck(1)\n1 Protein Sciences\ , Structure and Biophysics\, Discovery Sciences\, BioPharmaceuticals R&D\, AstraZeneca\, Gothenburg\, Sweden\; 2 Department of Chemistry and Molecul ar Biology\, University of Gothenburg\, Box 462\, SE-405 30 Gothenburg\, S weden. \n\nStructure-based drug design has played a pivotal role in pharma ceutical discovery for over thirty years\, leading to the development of n umerous approved therapeutics. Protein crystallography\, relying on data c ollection from large single crystals held at cryo temperature\, provides a highly optimised and effective workflow for generating structures of comp lexes between compounds and their target protein. Still\, this method requ ires manual manipulation of thousands of crystals per year for a company s uch as AstraZeneca. \nSerial crystallography presents an attractive altern ative\, offering potential improvements in throughput and automation by ci rcumventing labor-intensive crystal harvesting and facilitating streamline d sample preparation. Additionally\, the feasibility of room-temperature d ata collection can reduce the risk of structural artefacts introduced by c ryo-cooling. Nonetheless\, the adoption of serial crystallography in early drug discovery remains challenging due to constraints in speed\, protein and compound availability\, and workflow optimization.\nIn this study\, we demonstrate the development and implementation of an optimized serial cry stallography workflow tailored for drug discovery applications. Our approa ch enables the high-throughput screening of 384 fragments\, addressing som e key limitations while paving the way for broader application of serial c rystallography in pharmaceutical research.\n\nhttps://lindico453.srv.lu.se /event/583/contributions/1825/ LOCATION:LINXS at The Loop URL:https://lindico453.srv.lu.se/event/583/contributions/1825/ END:VEVENT BEGIN:VEVENT SUMMARY:Time-resolved serial crystallograhy to capture reaction intermedia tes of a glucuronyl esterase DTSTART;VALUE=DATE-TIME:20250922T154000Z DTEND;VALUE=DATE-TIME:20250922T155000Z DTSTAMP;VALUE=DATE-TIME:20260524T212503Z UID:indico-contribution-583-1833@lindico453.srv.lu.se DESCRIPTION:Speakers: Gabrielle Wehlander (University of Gothenburg)\nGluc uronyl esterases (GEs) from the carbohydrate esterase family 15 (CE15) are involved in degrading lignocellulosic biomass\, by catalyzing the hydroly sis of an esterbond connecting lignin and hemicellulose in the plant cell wall (1). In order to utilize biomass in biorefineries\, efficient methods are needed to separate cellulose\, hemicellulose and lignin. Studying GEs to better understand their reaction mechanism can aid in improving existi ng biological preteatment methods used in biorefineries to be able to make better use of this renewable energy source. The bacterial GE from Opitutu s terrae (OtCE15A) has previously been structurally determined at cryo-tem perature and a reaction mechanism for the acylation and deacylation reacti ons has been proposed (2\,3). Various glucuronate- and galacturonate ester s have been used as model substrates for the lignin-hemicellulose linkage\ , and the substrates have been soaked into the crystals. However\, attempt s to capture the binding of the substrates prior to hydrolysis of the este r bond have so far been unsuccessful\, but a covalent reaction intermediat e has been obtained using enzymes with mutations at the catalytic site. In attempts to capture the binding of substrates prior to hydrolysis and to determine reaction intermediates\, we have collected serial synchrotron X- ray crystallography (SSX) data at BioMAX (MAX IV\, Lund)\, and conducted i nitial time-resolved SSX experiment at P14.EH2 (T-REXX of PETRA III\, Hamb urg). We have obtained high resolution (1.7Å) SSX data of OtCE15A at BioM AX and observed binding of the cleaved substrate of benzyl glucuronoate af ter a soaking time of 5 minutes. For time-resolved SSX experiments at T-RE XX\, we have tested and are planning to use the hit-and-return (HARE) meth od (4).\n\nReferences\n1. Larsbrink\, Johan\, and Leila Lo Leggio. "Glucur onoyl esterases–enzymes to decouple lignin and carbohydrates and enable better utilization of renewable plant biomass." Essays in Biochemistry 67. 3 (2023): 493-503.\n2. Mazurkewich\, Scott\, et al. "Structural and bioche mical studies of the glucuronoyl esterase OtCE15A illuminate its interacti on with lignocellulosic components." Journal of Biological Chemistry 294.5 2 (2019): 19978-19987.\n3. Zong\, Zhiyou\, et al. "Mechanism and biomass a ssociation of glucuronoyl esterase: an α/β hydrolase with potential in b iomass conversion." Nature Communications 13.1 (2022): 1449.\n4. Schulz\, Eike C.\, et al. "The hit-and-return system enables efficient time-resolve d serial synchrotron crystallography." Nature methods 15.11 (2018): 901-90 4.\n\nAuthors: Gabrielle Wehlander\, Josefin Ridaeus\, Scott Mazurkewich\, Leila Lo Leggio\, Johan Larsbrink\, Gisela Brändén\n\nhttps://lindico45 3.srv.lu.se/event/583/contributions/1833/ LOCATION:LINXS at The Loop URL:https://lindico453.srv.lu.se/event/583/contributions/1833/ END:VEVENT BEGIN:VEVENT SUMMARY:Standard Sample Preparation and Characterization for Serial Crysta llography DTSTART;VALUE=DATE-TIME:20250922T171000Z DTEND;VALUE=DATE-TIME:20250922T172000Z DTSTAMP;VALUE=DATE-TIME:20260524T212503Z UID:indico-contribution-583-1842@lindico453.srv.lu.se DESCRIPTION:Speakers: Christina Schmidt (European XFEL)\nSerial crystallog raphy (SX) has been a revolutionary technique in structural biology for mo re than a decade\, providing insights into the structures and dynamics of biomolecules at room temperature. Using intense ultra-short X-ray pulses\, SX made the collection of diffraction data from micron-sized crystals pos sible\, avoiding radiation damage and allowing for the capture of transien t states and intermediates in biological reactions. The success of SX expe riments heavily relies on the quality of the sample.\nStandard samples in SX research provide critical roles. First\, the commissioning of new beaml ine devices needs well-characterized samples. By providing consistent and reproducible diffraction patterns\, the standard samples help in the calib ration of new devices and verification of their performance. Secondly\, th ey are required for the validation of experimental setups\, ensuring that all components\, from sample delivery systems to data acquisition software \, function correctly.\nIn this study\, we present detailed protocols for the preparation of lysozyme\, myoglobin\, iq-mEmerald\, and photoactive ye llow protein (PYP) crystals. These proteins were selected as standard samp les due to their robust crystallization properties and suitability for a w ide range of SX experiments. Through the optimization of existing protocol s\, we achieved high-quality crystal samples with improved yield\, specifi cally for SX applications.\n\n\nAuthors: Christina Schmidt\, Huijong Han\n \nhttps://lindico453.srv.lu.se/event/583/contributions/1842/ LOCATION:LINXS at The Loop URL:https://lindico453.srv.lu.se/event/583/contributions/1842/ END:VEVENT BEGIN:VEVENT SUMMARY:Time and dose resolved crystallography to control and capture redo x states in heme peroxidases DTSTART;VALUE=DATE-TIME:20250922T170000Z DTEND;VALUE=DATE-TIME:20250922T171000Z DTSTAMP;VALUE=DATE-TIME:20260524T212503Z UID:indico-contribution-583-1841@lindico453.srv.lu.se DESCRIPTION:Speakers: Michael Hough (Diamond Light Source)\nMetalloenzymes containing heme centres catalyse a wide range of reactions critical to li fe. Understanding the structure and electronic states of the heme centre a cross multiple functionally relevant states is essential to understand mec hanism. I will describe work using time resolved serial crystallography as well as the use of X-ray dose for the manipulation of heme iron oxidation states in dye decolourising peroxidases [1] using multiple\, complementar y\, serial crystallography and single-crystal spectroscopic approaches.\n\ nFixed target drop-on-chip\, tape drive droplet on demand and correlated s pectroscopies allow the formation of high valent Fe(IV) states to be chara cterised. X-ray Pump Probe serial femtosecond crystallography (SFX) togeth er with dose-resolved serial synchrotron crystallography (SSX) allowed the peroxidases to be driven between multiple iron oxidation states that can be spectroscopically validated. Intriguingly\, the formation and dose resp onse of the Fe(IV)-O state is highly variable between the chemically ident ical heme groups of the homo-oligomeric proteins highlighting the importan ce of understanding the effect of the crystalline lattice on observed chan ges in time- and dose-resolved crystallography experiments.\n\nLucic\, M. et al (2021) Aspartate or arginine? Validated redox state X-ray structures elucidate mechanistic subtleties of FeIV = O formation in bacterial d ye-decolorizing peroxidases. JBIC 27 (7)\, 743-761.\n\n\nCo-authors: Robin Owen\, Jonathan Worrall\, Marina Rozman\, Lewis Williams\, Danny Axford\, Allen Orville\, Pierre Aller\, Jan Kern\, Jos Kamps\n\nhttps://lindico453 .srv.lu.se/event/583/contributions/1841/ LOCATION:LINXS at The Loop URL:https://lindico453.srv.lu.se/event/583/contributions/1841/ END:VEVENT BEGIN:VEVENT SUMMARY:Experimental estimation of copper-ligand length precision in a mod el fungal LPMO under redox cycling and saccharide binding DTSTART;VALUE=DATE-TIME:20250922T165000Z DTEND;VALUE=DATE-TIME:20250922T170000Z DTSTAMP;VALUE=DATE-TIME:20260524T212503Z UID:indico-contribution-583-1840@lindico453.srv.lu.se DESCRIPTION:Speakers: Zhiyu Huang (University of Copenhagen)\nLytic polysa ccharide monooxygenases (LPMOs) are copper-dependent enzymes that degrade polysaccharides oxidatively\, with applications in second-generation bioet hanol production and as virulence factors in certain pathogens [1] [2]. Th ey have been reclassified within the CAZy database into auxiliary activity families AA9–AA11 and AA13–AA17 [3]\, and their active-site histidine brace is highly conserved. The catalytic mechanism of LPMOs is complex\, with the priming reaction step requiring the reduction of Cu(II) to Cu(I). Since the geometric changes associated with redox cycling—whether chemi cally induced or triggered by photoreduction—can be subtle\, the accurat e determination of bond lengths and angles is essential [4]. In this study \, LsAA9A from Lentinus similis was used as a model system under various e xperimental conditions. Analysis of the LsAA9A_Ec and LsAA9A_Ec_Cell3 stru ctures collected under low-dose conditions showed that\, compared with oth er Cu-coordination distances\, only the Tyr–Cu bond exhibited a statisti cally significant change (p = 0.00094 in two tailed t-test). These finding s confirm that saccharide substrate binding consistently shortens the Tyr –Cu distance in LsAA9A_Ec in the Cu²⁺ state\, with a measured reducti on of 0.21 Å. Furthermore\, experiments on LsAA9A_Ec_Asc and LsAA9A_Ec_As c_Cell3\, conducted under both low- and high-dose conditions\, where Cu² ⁺ can be reduced to Cu⁺ by X-ray exposure\, as well as at room tempera ture\, further probed structural responses to varying redox and experiment al regimes. These findings advance the mechanistic understanding of LPMOs and offer a framework for probing subtle geometric changes in metalloenzym es.\n\nAuthors: Zhiyu Huang\, Jie Nan\, Monika Bjelcic\, Leila Lo Leggio\n \nhttps://lindico453.srv.lu.se/event/583/contributions/1840/ LOCATION:LINXS at The Loop URL:https://lindico453.srv.lu.se/event/583/contributions/1840/ END:VEVENT BEGIN:VEVENT SUMMARY:Serial X – Simple Solutions for Serial Synchrotron Crystallograp hy DTSTART;VALUE=DATE-TIME:20250922T164000Z DTEND;VALUE=DATE-TIME:20250922T165000Z DTSTAMP;VALUE=DATE-TIME:20260524T212503Z UID:indico-contribution-583-1844@lindico453.srv.lu.se DESCRIPTION:Speakers: Yanyan Chen (Serial X AB)\nSerial X aims to expand o f the use of serial crystallography across the academic and industrial lif e-science community by introducing low cost\, flexible and easy to use sol utions to the problem of sample delivery in serial crystallography experim ents at synchrotron radiation facilities. These solutions were invented in ProtonPump\, the ERC Advanced Grant awarded to Richard Neutze\, which app lies time-resolved serial crystallography to observe structural changes in the enzyme cytochrome c oxidase. Serial crystallography is going through a period of rapid development and the number of scientific users of this m ethod has the potential to grow by up-to two orders of magnitude. This pot ential will never be realized if the current lack of standardization in sa mple delivery continues\, which is prohibitively expensive and is often un reliable. Serial-X will solve this problem by developing and bringing to m arket standardized\, low-cost\, flexible and easy-to-use solutions for sam ple delivery in serial crystallography studies at synchrotron radiation so urces. With the support of ERC proof-of-concept grant to Richad Neutze\, S erial X has developed low-cost products supporting both flow cell1 and fix ed target2 approaches to serial crystallography that are mounted upon stan dard magnets used at all conventional macromolecular X-ray crystallography synchrotron-based beamlines. This will remove the greatest obstacle curre ntly preventing scientists from using serial crystallography for their own research or for structure based drug-design within a pharmaceutical drug discovery context.\n\n\nAuthors: Yanyan Chen\, Swagatha Ghosh\, Gisela Br ändén\, Richard Neutze\n\nhttps://lindico453.srv.lu.se/event/583/contrib utions/1844/ LOCATION:LINXS at The Loop URL:https://lindico453.srv.lu.se/event/583/contributions/1844/ END:VEVENT BEGIN:VEVENT SUMMARY:Reducing data volume with X-ray Laue diffraction DTSTART;VALUE=DATE-TIME:20250922T163000Z DTEND;VALUE=DATE-TIME:20250922T164000Z DTSTAMP;VALUE=DATE-TIME:20260524T212503Z UID:indico-contribution-583-1839@lindico453.srv.lu.se DESCRIPTION:Speakers: Kah Chee Pow (Spallation Neutron Source Science Cent er)\nThe X-ray Laue diffraction captures the crystal diffraction from whit e beam and can contain significant amount of structural information in com parison to monochromatic diffraction. In time-resolved study\, the serial femtosecond crystallography (SFX) and serial synchrotron crystallography ( SSX) are the mainstream approaches with the caveat of generating large dat a volume. Laue diffraction has great potential in mitigating the data volu me challenge and has recently regain the interest by the community. Here\, we share the preliminary works conducted at BL03HB Laue Micro-diffraction Beamline at Shanghai Synchrotron Radiation Facility (SSRF) to compare wit h conventional method. We crystallized an apo stilbene synthase (STS) with inherent loop conformation duality and collected diffraction data via (i) single-crystal\, cryogenic\, rotation approach with monochromatic beam\, (ii) single-crystal\, ambient\, helical approach with white beam\, and (ii i) multi-crystal\, ambient\, single-frame approach with white beam. Compar e to monochromatic diffraction\, Laue diffraction dataset show increased c ontent of structural information per image but with poor statistical metri cs upon merging. Nevertheless\, the solved structure from Laue diffraction dataset can resolve the duality of the loop conformation and reveal the l oop conformational preference at productive state at ambient temperature. In short\, Laue diffraction is a promising approach to explore in mitigati ng the data volume challenge.\n\nAuthor: Kah Chee Pow\nCo-author: Quan Hao \n\nhttps://lindico453.srv.lu.se/event/583/contributions/1839/ LOCATION:LINXS at The Loop URL:https://lindico453.srv.lu.se/event/583/contributions/1839/ END:VEVENT BEGIN:VEVENT SUMMARY:Structural studies of the human drug-metabolising protein CYP3A4 DTSTART;VALUE=DATE-TIME:20250922T161000Z DTEND;VALUE=DATE-TIME:20250922T162000Z DTSTAMP;VALUE=DATE-TIME:20260524T212503Z UID:indico-contribution-583-1838@lindico453.srv.lu.se DESCRIPTION:Speakers: Johan Glerup (University of Gothenburg)\nA highly fl exible protein with an active site that changes its volume to fit a wide v ariety of ligands. A lid made of loops changes conformation based on ligan d-size. Inhibition of this enzyme stops metabolism of drugs. This is an au tomatic disqualification of a drug candidate. Room temperature SSX shows b etter definition of some flexible loops\, even at worse resolution. SSX da ta collection at tens of kilo Gray produces a similar active-site to our X FEL structure. I present an internal distance matrix analysis of a subset of PDB CYP3A4 structures to determine that crystal form\, resolution and t o some extent ligand-size dominates the clustering of global similarity wi th little difference caused by temperature. The protein crystalises as a m onomer in the ASU but SAX\, cryo-EM and SEC-MALS shows a homo-tetramer in solution bringing it into the perfect size range for cryo-EM. I present in itial data for the volume of the tetramer solved through single particle a nalysis at SciLifeLab Solna.\n\n\nCo-authors: Owens Uwangue\, Gisela Brand en\, Monika Bjelcic\n\nhttps://lindico453.srv.lu.se/event/583/contribution s/1838/ LOCATION:LINXS at The Loop URL:https://lindico453.srv.lu.se/event/583/contributions/1838/ END:VEVENT BEGIN:VEVENT SUMMARY:Development of new data processing methods for serial time-resolve d crystallography DTSTART;VALUE=DATE-TIME:20250922T160000Z DTEND;VALUE=DATE-TIME:20250922T161000Z DTSTAMP;VALUE=DATE-TIME:20260524T212503Z UID:indico-contribution-583-1837@lindico453.srv.lu.se DESCRIPTION:Speakers: Rachel Tang (Diamond Light Source)\nTo obtain the hi ghest quality electron density maps\, from a time-resolved serial crystall ography experiment\, it is crucial to accurately group the observed intens ities according to the structure of the protein that generated those inten sities. In an ideal world\, each unique protein crystal structure would gi ve rise to a set of well defined intensities\, which would enable the inte nsities to be grouped easily. Unfortunately\, due to many real world limit ations\, this is not the case: 1) The crystal is an average of many protei n structures 2) Experimental effects such as unequal soaking\, unequal las er/ x-ray exposure etc. 3) A range of crystal sizes causing inconsistent i ntensities which require rescaling. These effects give rise to uncertainty in the observed intensity values. Furthermore\, only a minor proportion o f hkls are affected by the change in protein structure\, and of those hkls \, the intensity changes are often subtle.\nDespite all the challenges\, a statistically rigorous approach is required to accurately group observed intensities according to the structure of the protein that generated those intensities. One approach utilises Bayesian statistics\, a probability ba sed approached used in many areas such as weather forecasting\, econometri cs and natural language processing. In principle\, some Bayesian methods s uch as naive Bayes and maximum likelihood estimator is able to assess the probability a set of intensities were derived from a protein structure. In practice\, the intensities have high uncertainty values and there are man y nuances as to how this data processing pipeline should be set up\, somet imes sacrificing flexibility for increased confidence in the results.\n\nC o-authors: James Beilsten-Edmands\, Mike Hough\, Graeme Winter\n\nhttps:// lindico453.srv.lu.se/event/583/contributions/1837/ LOCATION:LINXS at The Loop URL:https://lindico453.srv.lu.se/event/583/contributions/1837/ END:VEVENT BEGIN:VEVENT SUMMARY:Comprehensive Support for Serial Crystallography at the European X FEL DTSTART;VALUE=DATE-TIME:20250922T155000Z DTEND;VALUE=DATE-TIME:20250922T160000Z DTSTAMP;VALUE=DATE-TIME:20260524T212503Z UID:indico-contribution-583-1836@lindico453.srv.lu.se DESCRIPTION:Speakers: Huijong Han (European XFEL)\nSerial crystallography has opened new possibilities for determining the structures of biological macromolecules using microcrystals. At the European XFEL\, the Sample Envi ronment and Characterization (SEC) group\, operating the XBI biology labor atory\, provides essential support for users at every stage of the serial crystallography workflow\, from initial sample preparation to advanced cha racterization and sample delivery.\nThe XBI laboratory offers state-of-the -art facilities where users can prepare and characterize their samples bef ore experiments. Through hands-on support and guidance\, we help users opt imizing their samples to make efficient use of valuable beamtime. The SEC group provides sample delivery methods and supports users in optimizing th eir own systems.\nBy combining expertise in sample delivery and advanced c haracterization\, the SEC group at European XFEL supports the structural b iology community to achieve innovative results in serial crystallography. Our ongoing developments in standard samples\, support infrastructure\, an d efficient sample delivery methods continue to lower access barriers\, fo ster collaboration\, and broaden the access to serial crystallography.\n\n https://lindico453.srv.lu.se/event/583/contributions/1836/ LOCATION:LINXS at The Loop URL:https://lindico453.srv.lu.se/event/583/contributions/1836/ END:VEVENT BEGIN:VEVENT SUMMARY:In crystallo study of the reaction mechanism in a family B DNA pol ymerase DTSTART;VALUE=DATE-TIME:20250922T153000Z DTEND;VALUE=DATE-TIME:20250922T154000Z DTSTAMP;VALUE=DATE-TIME:20260524T212503Z UID:indico-contribution-583-1835@lindico453.srv.lu.se DESCRIPTION:Speakers: Vimal Parkash (Umeå University)\nReaction intermedi ates during DNA synthesis have been studied in detail using time-resolved\ nX-ray crystallography for translesion and repair DNA polymerases. Contrar y to the originally\nproposed two-metal-ion mechanism\, a third metal ion was identified between the finger\ndomain and the α- and β-phosphates of the incoming nucleotide. This third metal ion was\nsuggested to either pa rticipate in catalysis or stabilize product formation. To investigate this \nfurther in a replicative polymerase\, we conducted time-resolved X-ray c rystallography with\nDNA Polymerase epsilon\, which synthesizes DNA at a m uch faster rate—10x to 100x higher\nthan family Y and X polymerases. Sur prisingly\, no metal ion was observed between the finger\ndomain and the α- and β-phosphates of the incoming nucleotide in any of the solved stru ctures\nwith Pol epsilon. Instead\, our biochemical and structural data su pport the original two-metal\nmechanism. In addition\, we discovered that the 3’-OH group releases a proton\, which is\nchanneled via structural w aters to a basic residue in the Palm domain. After forming a new\nbond wit h the incoming nucleotide\, an acidic residue in the finger domain protona tes the\nreleased pyrophosphate\, stabilizing the product. In summary\, it seems that metal A’s role is\nto lower the pKa of the 3’-OH group\, f ollowed by specific residues in Pol epsilon donating or\nreceiving a proto n to catalyze this acid-base reaction.\n\nCo-author: Erik Johansson\n\nhtt ps://lindico453.srv.lu.se/event/583/contributions/1835/ LOCATION:LINXS at The Loop URL:https://lindico453.srv.lu.se/event/583/contributions/1835/ END:VEVENT BEGIN:VEVENT SUMMARY:Wrap-up DTSTART;VALUE=DATE-TIME:20250924T094500Z DTEND;VALUE=DATE-TIME:20250924T100000Z DTSTAMP;VALUE=DATE-TIME:20260524T212503Z UID:indico-contribution-583-1861@lindico453.srv.lu.se DESCRIPTION:https://lindico453.srv.lu.se/event/583/contributions/1861/ LOCATION:LINXS at The Loop URL:https://lindico453.srv.lu.se/event/583/contributions/1861/ END:VEVENT BEGIN:VEVENT SUMMARY:Serial Microsecond Crystallography with the ESRF Extremely Brillia nt Source DTSTART;VALUE=DATE-TIME:20250924T091500Z DTEND;VALUE=DATE-TIME:20250924T094500Z DTSTAMP;VALUE=DATE-TIME:20260524T212503Z UID:indico-contribution-583-1860@lindico453.srv.lu.se DESCRIPTION:Speakers: Daniele de Sanctis (ESRF - The European Synchrotron) \nTime-resolved serial crystallography (TR-SX) is a leading technique for capturing biological processes as molecular movies on extremely fast times cales—fulfilling a long-standing goal in structural biology. The method involves delivering microcrystals into a powerful\, pulsed X-ray beam to c ollect individual diffraction patterns from each crystal. By compiling tho usands of these patterns\, researchers can reconstruct an electron density map. This technique\, known as serial crystallography\, allows for the ob servation of structural changes. When ultrashort X-ray pulses are used\, S X can track time-dependent conformational changes\, enabling scientists to visualize proteins in motion. TR-SX experiments are typically synchronize d with a specific stimulus that initiates the biological activity under in vestigation. The advent of diffraction limited storage rings - the so-call ed 4th generation synchrotrons - have permitted the conception and built i nstruments that overcomes the limits of traditional microfocus beamlines. These new beamlines aim to exploit X-ray pulses down to microsecond time r esolution\, becoming an invaluable tool for room temperature and time-reso lved studies that complements the capabilities of Free Electron Laser sour ces. ID29 at the European Synchrotron Radiation Facility is one of the fir st examples of this new generation of beamlines (Orlans et al. 2025). The unique combination of microsecond exposure times\, advanced beam propertie s\, and a flexible sample environment enables the collection of high-quali ty\, complete data—even from exceptionally small amounts of crystalline material. This is applied in combination with external stimuli to activate or induce structural changes that could be observed in the microsecond ti me regime. This approach is particularly successful for the study of enzym atic reaction or ligand binding\, thus prominently interesting for the who le structural biology community\, while future developments will be crucia l to strengthen the application of the methods to structural based drug de sign.\n\nOrlans\, J. (2025). Advancing macromolecular structure determinat ion with microsecond X-ray pulses at a 4th generation synchrotron. Communi cations Chemistry\, 8(1)\, 1–12. https://doi.org/10.1038/s42004-024-0140 4-y\n\nhttps://lindico453.srv.lu.se/event/583/contributions/1860/ LOCATION:LINXS at The Loop URL:https://lindico453.srv.lu.se/event/583/contributions/1860/ END:VEVENT BEGIN:VEVENT SUMMARY:Serial Techniques for Weakly Scattering\, In-Situ\, and Dynamic Sy stems DTSTART;VALUE=DATE-TIME:20250924T084500Z DTEND;VALUE=DATE-TIME:20250924T091500Z DTSTAMP;VALUE=DATE-TIME:20260524T212503Z UID:indico-contribution-583-1859@lindico453.srv.lu.se DESCRIPTION:Speakers: Mark Warren (Diamond Light Source)\nhttps://lindico4 53.srv.lu.se/event/583/contributions/1859/ LOCATION:LINXS at The Loop URL:https://lindico453.srv.lu.se/event/583/contributions/1859/ END:VEVENT BEGIN:VEVENT SUMMARY:Turning Up the Heat on Dynamic Proteins with Temperature-Jump X-ra y Crystallography DTSTART;VALUE=DATE-TIME:20250924T081500Z DTEND;VALUE=DATE-TIME:20250924T084500Z DTSTAMP;VALUE=DATE-TIME:20260524T212503Z UID:indico-contribution-583-1858@lindico453.srv.lu.se DESCRIPTION:Speakers: Michael Thompson (University of California\, Merced) \nProtein dynamics are critical for function\, but it remains challenging to understand\, in atomic detail\, how a molecule’s biological activity is enabled by the physical coupling of its conformational fluctuations acr oss varied length and time scales. Time-dependent X-ray crystallographic m easurements of molecular structure can overcome some of the limitations of traditional structural biology and yield deep insight into protein confor mational landscapes\, but it remains challenging to initiate synchronous c onformational changes in crystallized macromolecules\, which is a requirem ent for such experiments. I will describe how observations from multi-temp erature structural measurements motivated the development of temperature-j ump (T-jump) crystallography\, and summarize the results of our early T-ju mp experiments on the model enzyme lysozyme. I will also discuss ongoing e fforts to democratize these experiments and apply them to increasingly com plex biological systems\, including the metalloenzyme soybean lipoxygenase \, whose catalytic mechanism involves a rate-limiting hydrogen tunneling s tep that is coupled to motion of the protein scaffold.\n\nhttps://lindico4 53.srv.lu.se/event/583/contributions/1858/ LOCATION:LINXS at The Loop URL:https://lindico453.srv.lu.se/event/583/contributions/1858/ END:VEVENT BEGIN:VEVENT SUMMARY:Time-resolved macromolecular crystallography studies of AmpCEC usi ng synchrotron and XFEL radiation DTSTART;VALUE=DATE-TIME:20250924T073000Z DTEND;VALUE=DATE-TIME:20250924T074500Z DTSTAMP;VALUE=DATE-TIME:20260524T212503Z UID:indico-contribution-583-1857@lindico453.srv.lu.se DESCRIPTION:Speakers: Emily Freeman (University of Oxford)\nThe β-lactama se enzymes degrade β-lactam antibiotics\, exemplified by penicillin. As s uch\, the families of metallo- and serine β-lactamases are responsible fo r a major antimicrobial resistance mechanism in many clinically relevant s pecies of Gram-negative bacteria\, including Escherichia coli\, Klebsiella pneumoniae and Pseudomonas aeruginosa. To preserve the antimicrobial acti vity of β-lactam antibiotics\, inhibitors of β-lactamases can be used in combination with a β-lactam antibiotic during treatment of an antimicrob ial resistant infection. However\, these inhibitors often have a narrow sp ectrum of activity against β-lactamases\, and other bacterial mechanisms of resistance against them. To further development of novel β-lactamase i nhibitors\, we apply time-resolved serial femtosecond and synchrotron crys tallography (tr-SF/SX) techniques to investigate the acylation of the β-l actamase AmpC from Escherichia coli by avibactam\, a clinically approved β-lactamase inhibitor. Previously\, tr-SF/SX data gathered by using a “ drop-on-demand” sample delivery system revealed that covalent binding of avibactam to the AmpC active site occurred in a time frame quicker than 2 00 ms and as such pre-acylated time-resolved structures could not be obtai ned using this system. In the interest of capturing earlier time points us ing this enzymatic system\, we have turned our focus to using a “drop-on -chip” fixed target sample delivery system\, addressing contamination is sues and implementing robust controls in our setup for accurate data colle ction. Currently\, we are employing a drop-on-chip fixed target sample del ivery system to access time points >1 ms at XFELs and >10 ms at synchrotro ns such as Diamond Light Source.\n\nAuthors:\nEmily Freeman\, Jos Kamps\, Pierre Aller\, Christopher Schofield\, Allen Orville\n\nhttps://lindico453 .srv.lu.se/event/583/contributions/1857/ LOCATION:LINXS at The Loop URL:https://lindico453.srv.lu.se/event/583/contributions/1857/ END:VEVENT BEGIN:VEVENT SUMMARY:Investigating enzyme mechanisms by multidimensional crystallograph y DTSTART;VALUE=DATE-TIME:20250924T070000Z DTEND;VALUE=DATE-TIME:20250924T073000Z DTSTAMP;VALUE=DATE-TIME:20260524T212503Z UID:indico-contribution-583-1856@lindico453.srv.lu.se DESCRIPTION:Speakers: Pedram Mehrabi (University of Hamburg)\nFunctional c haracterization of proteins requires linking structure and dynamics\, but traditional X-ray crystallography provides only static snapshots. Serial t ime-resolved crystallography enables direct visualization of structural ch anges over time\, including internal motions and solvent interactions. We developed fixed-target approaches such as “Hit And REturn” (HARE) and reaction initiation strategies using piezo droplet injectors. The “Liqui d Application Method for time-resolved Analysis” (LAMA) further broadens applicability to systems not triggered by light. In addition\, environmen tal control allows temperature variation from 7 °C to 70 °C\, enabling m ulti-dimensional experiments. These advances permit direct observation of ligand binding\, intermediates\, and conformational changes\, as demonstra ted by tracking glucose-to-fructose conversion in Xylose isomerase across both temperature and time. Together\, these methods expand the toolkit for time-resolved crystallography\, opening the way to mechanistic insights i nto enzyme dynamics\, allostery\, and solvent networks.\n\nhttps://lindico 453.srv.lu.se/event/583/contributions/1856/ LOCATION:LINXS at The Loop URL:https://lindico453.srv.lu.se/event/583/contributions/1856/ END:VEVENT BEGIN:VEVENT SUMMARY:MAX IV visit DTSTART;VALUE=DATE-TIME:20250923T134500Z DTEND;VALUE=DATE-TIME:20250923T150000Z DTSTAMP;VALUE=DATE-TIME:20260524T212503Z UID:indico-contribution-583-1855@lindico453.srv.lu.se DESCRIPTION:https://lindico453.srv.lu.se/event/583/contributions/1855/ LOCATION:LINXS at The Loop URL:https://lindico453.srv.lu.se/event/583/contributions/1855/ END:VEVENT BEGIN:VEVENT SUMMARY:Panel discussion DTSTART;VALUE=DATE-TIME:20250923T123000Z DTEND;VALUE=DATE-TIME:20250923T130000Z DTSTAMP;VALUE=DATE-TIME:20260524T212503Z UID:indico-contribution-583-1854@lindico453.srv.lu.se DESCRIPTION:Speakers: John Beale (Paul Scherrer Institute)\nModerator: Dr. John Beale\, Paul Scherrer Institute\n\nProf. Guillermo Montoya\, Univers ity of Copenhagen\nProf. Gisela Brändén\, University of Gothenburg\nDr. Judith Juanhuix\, ALBA-CELLS\n\nhttps://lindico453.srv.lu.se/event/583/con tributions/1854/ LOCATION:LINXS at The Loop URL:https://lindico453.srv.lu.se/event/583/contributions/1854/ END:VEVENT BEGIN:VEVENT SUMMARY:Quantum refinement used for time-resolved crystallography DTSTART;VALUE=DATE-TIME:20250923T094500Z DTEND;VALUE=DATE-TIME:20250923T100000Z DTSTAMP;VALUE=DATE-TIME:20260524T212503Z UID:indico-contribution-583-1850@lindico453.srv.lu.se DESCRIPTION:Speakers: Gayathri Yuvaraj (Lund University)\nIn standard crys tallographic refinement of proteins\, the experimental data are normally n ot enough to unambiguously decide the positions of all atoms. Therefore\, the crystallographic data are supplemented by a set of empirical restraint s that ensure that bond lengths and angles make chemical sense. To obtain more accurate results\, we have suggested that this potential can be repla ced by more accurate quantum-mechanical (QM) calculations for a small\, bu t interesting part of the protein\, giving the method of quantum refinemen t.1 Our group has shown that quantum refinement can locally improve crysta l structures\,2 decide protonation state of metal-bound ligands\,3–6 oxi dation state of metal sites\,7\,8 detect photoreduction of metal ions7\,9 and solve scientific problems regarding what is really is seen in crystal structures.9–11 Several other groups have implemented this and similar a pproaches.12 We investigate how quantum refinement can be used for time-re solved crystallography. In time-resolved crystallography\, the obtained el ectron-density maps will typically involve a mixture of several states (un reacted state\, intermediates and products). Therefore\, the structures wi ll heavily depend on the empirical potential and the expectations of the c rystallographer. The QM calculations will give more accurate results\, esp ecially if there are intermediates with unusual (e.g. twisted) structures or if metal sites are involved (which are hard to describe with general re straints). Moreover\, we will couple the structural interpretations with e xpectations from kinetic models of the studied reaction. I will present so me preliminary applications on cytochrome c oxidase\, xylose isomerase and bacteriorhodopsin.\n\nReferences\n1. U. Ryde\, L. Olsen\, K. Nilsson\, 20 02\, J. Comput. Chem. 23\, 1058.\n2. U. Ryde\, K. Nilsson J. Am. Chem. Soc . 2003\, 125\, 14232.\n3. K. Nilsson\, U. Ryde\, J. Inorg. Biochem.\, 2004 \, 98\, 1539\n4. L. Cao\, O. Caldararu\, U. Ryde\, J. Phys. Chem B\, 2017\ , 121\, 8242.\n5. L. Cao\, O. Caldararu\, U. Ryde\, J. Chem. Theory Comput .\, 2018\, 14\, 6653.\n6. O. Caldararu\, M. Feldt\, D. Cioloboc\, M.-C.van Severen\, K. Starke\, E. Nordlander\, et al. Sci. Rep. 2018\, 8\, 4684\n7 . L. Rulíšek\, U. Ryde\, J. Phys. Chem. B\, 2006\, 110\, 11511\n8. L. Ca o\, Börner\, M. C.\, Bergmann\, J.\, Caldararu\, O. & U. Ryde\, Inorg. Ch em. 2019\, 58\, 9672.\n9. P. Söderhjelm\, U. Ryde\, J. Mol. Struct. Theoc hem\, 2006\, 770\, 199\n10. L. Cao\, O. Caldararu\, A. C. Rosenzweig\, U. Ryde\, 2018\, Angew. Chem. Int. Ed.\, 57\,162.\n11. J. Bergmann\, E. Oksan en & U. Ryde\, J. Biol. Inorg. Chem. 2021\, 26\, 341.\n12. J. Bergmann\, E . Oksanen\, U. Ryde\, Curr. Opin. Struct. Biol. 2022\, 72\, 18.\n\n\nAutho rs:\nGayathri Yuvaraj\, Ulf Ryde\nCo-authors:\nKristoffer Lundgren\, Esko Oksanen\n\nhttps://lindico453.srv.lu.se/event/583/contributions/1850/ LOCATION:LINXS at The Loop URL:https://lindico453.srv.lu.se/event/583/contributions/1850/ END:VEVENT BEGIN:VEVENT SUMMARY:Modeling X-ray-Induced Heating at 4th-Generation MX Beamlines DTSTART;VALUE=DATE-TIME:20250923T091500Z DTEND;VALUE=DATE-TIME:20250923T094500Z DTSTAMP;VALUE=DATE-TIME:20260524T212503Z UID:indico-contribution-583-1849@lindico453.srv.lu.se DESCRIPTION:Speakers: Michal Kepa (Paul Scherrer Institute)\nThe transitio n from 3rd to 4th generation synchrotrons—featuring diffraction-limited storage rings—has significantly expanded future possibilities for macrom olecular crystallography (MX). These upgrades\, characterized by reduced s ource divergence and increased electron bucket capacity\, have boosted bri lliance by up to two orders of magnitude. As a result\, next-generation MX beamlines\, such as ID29 (ESRF-EBS)\, BioCARS (APS-U)\, and MicroMAX (MAX IV)\, now deliver fluxes approaching 10^15 ph/s. These extremely high flu x (EHF) beamlines are increasingly optimized for time-resolved MX\, aiming for microsecond-scale resolution.\n\nOperating in previously unexplored d ose regimes (>50 GGy/s) raises new challenges. In my presentation\, I woul d like to share our study that focuses on beam-induced heating in microcry stals (<25 µm) exposed to EHF conditions. Thermal modeling indicates that such dose rates may cause significant temperature rises\, potentially imp acting data quality. Mitigation strategies include using top-hat beam prof iles and increasing both beam and crystal sizes to distribute dose more ev enly. The proposed model serves as a tool to support experimental design a nd optimize conditions for high-flux time-resolved MX. This of critical im portance for future multidimensional X-ray protein crystallography\, espec ially in enzymology\, where temperature is a key experimental variable. As such they will require precise temperature measurements\, as even a small change in temperature can affect the catalytic activity of an enzyme.\n\n \nAuthors: Michal Kepa\, John Beale\, Martin Appleby\n\nhttps://lindico453 .srv.lu.se/event/583/contributions/1849/ LOCATION:LINXS at The Loop URL:https://lindico453.srv.lu.se/event/583/contributions/1849/ END:VEVENT BEGIN:VEVENT SUMMARY:GPCRs as Targets for Serial Crystallography DTSTART;VALUE=DATE-TIME:20250923T080000Z DTEND;VALUE=DATE-TIME:20250923T081500Z DTSTAMP;VALUE=DATE-TIME:20260524T212503Z UID:indico-contribution-583-1847@lindico453.srv.lu.se DESCRIPTION:Speakers: Hannah Glover (leadXpro)\nUsing time-resolved serial crystallography to observe structural snapshots of protein dynamics at hi gh resolution is a method that is becoming gradually more commonplace. Adv ancements in method development for this technique have allowed a wider ra nge of proteins to be studied\; looking at processes spanning endogenous p hotoresponses\, enzyme kinetics and ligand binding. G protein-coupled rece ptors represent a pharmacologically relevant superfamily of proteins that are interesting targets for study with time-resolved serial crystallograph y. Data from time-resolved serial crystallography has the potential to enh ance the drug design process by revealing protein transitional states that can be either targeted or used to provide information about protein flexi bility. Our goal is to study the inherent dynamics of GPCRs critical for r eceptor function and to use this information to develop more targeted liga nds. Here\, we present the results of time-resolved serial crystallography experiments conducted at MaxIV and the SLS on the human A2a receptor. Thr ough synthetic photoswitches\, based on the marketed drug istradefylline f or the treatment of Parkinson’s disease\, light is used as a trigger to investigate the dynamics associated with ligand dissociation from the rece ptor orthosteric binding pocket. Our time-resolved data highlights key str uctural features involved in the transition upon ligand photoswitching. Th is includes the rearrangement of extracellular loops 2 and 3 that form a l id over the binding pocket\, which has been shown by molecular dynamic sim ulations\, crystal structures and kinetic analyses to be crucial for ligan d dissociation and long target resident time. Additionally\, lessons learn ed from this investigation\, in terms of experimental design and sample pr eparation\, can be applied to future projects using GPCRs as targets for s erial crystallography. Helping to lower the barrier of entry to time-resol ved serial crystallography and ultimately leading to more rationally desig ned drugs.\n\n\nCo-authors:\nTorben Saßmannshausen\, Quentin Bertrand\, M atilde Trabuco\, Chavdar Slavov\, Arianna Bacchin\, Fabio Andres\, Yasushi Kondo\, Robin Stipp\, Maximilian Wranik\, Georgii Khusainov\, Melissa Car rillo\, Demet Kekilli\, Jie Nan\, Ana Gonzalez\, Robert Cheng\, Werner Nei dhart\, Tobias Weinert\, Filip Leonarski\, Florian Dworkowski\, Michal Kep a\, Josef Wachtveitl\, Michael Hennig\, Joerg Standfuss\n\nhttps://lindico 453.srv.lu.se/event/583/contributions/1847/ LOCATION:LINXS at The Loop URL:https://lindico453.srv.lu.se/event/583/contributions/1847/ END:VEVENT BEGIN:VEVENT SUMMARY:Serial crystallography at the beamline P11\, PETRA III DTSTART;VALUE=DATE-TIME:20250922T131000Z DTEND;VALUE=DATE-TIME:20250922T132500Z DTSTAMP;VALUE=DATE-TIME:20260524T212503Z UID:indico-contribution-583-1829@lindico453.srv.lu.se DESCRIPTION:Speakers: Guillaume Pompidor (DESY)\nP11 at PETRA III (DESY\, Hamburg) is a high-throughput instrument for macromolecular crys-\ntallogr aphy [1]. The beamline has tuneable photon energy between 5.5 - 28 keV and and beam\nsizes from 200 x 200 μm to 4 x 9 μm2 can be used with a maxim um photon flux of 1x1013 ph/s at\n12 keV energy. Equipped with a fast dete ctor Eiger2 X 16M as the stationary detector (max 133 Hz)\,\nthis high flu x instrument is perfect for serial synchrotron crystallography (SSX).\nFor SSX\, the sample delivery is achieved through various types of solid supp orts and the TapeDrive\nsetup\, which allows time-resolved room temperatur e experiments by the mix-and-diffuse method\n[2\, 3]\, and has been develo ped at P11 along with the real-time autoprocessing with CrystFEL [4\,\n5]. In this method\, the samples are delivered on to a continuously drawn pol yimide tape through\na 3D-printed microfluidic nozzle with two channels ( ø 150 μm each)\, one for the crystal slurry and\nsecond\, for example\, for mixing the crystals with a ligand or adjusting their pH. The mixing ti me\ncan be modified by the speed of the tape and the distance of the nozzl e from the X-ray focus point\n(delay times of 50 ms – 180 s).\nIn order to mitigate possible radiation damage\, the exposure time per frame can be reduced from\n7.5 ms instead of 3.5 ms with a chopper wheel. SSX experime nts are currently controlled through\na separate graphical user interface\ , and online data analysis is available for real-time evaluation\nand inde xing via OnDA Monitor [6].\nWe will present the TapeDrive setup used for S SX\, the real time data processing implemented as\nwell as several scienti fic examples exploiting the capabilities of the setup (time-resolved\, mix -and-\ndiffuse experiments\, temperature control…).\n[1] Burkhardt\, A.\ , Pakendorf\, T.\, Reime\, B.\, Meyer\, J.\, Fischer\, P.\, Stübe\, N.\, Panneerselvam\, S.\,\nLorbeer\, O.\, Stachnik\, K.\, Warmer\, M.\, Rödig\ , P.\, Göries\, D. & Meents\, A. (2016). Eur. Phys. J. Plus\n131\, 56.\n[ 2] Beyerlein\, K. R.\, Dierksmeyer\, D.\, Mariani\, V.\, Kuhn\, M.\, Sarro u\, I.\, Ottaviano\, A.\, Awel\, S.\,\nKnoska\, J.\, Fuglerud\, S.\, Jöns son\, O.\, Stern\, S.\, Wiedorn\, M. O.\, Yefanov\, O.\, Adriano\, L.\, Be an\, R.\,\nBurkhardt\, A.\, Fischer\, P.\, Heymann\, M.\, Horke\, D. A.\, Jungnickel\, K. E. J.\, Kovaleva\, E.\, Lorbeer\, O.\,\nMetz\, M.\, Meyer\ , J.\, Morgan\, A.\, Pande\, K.\, Panneerselvam\, S.\, Seuring\, C.\, Tols tikova\, A.\, Lieske\,\nJ.\, Aplin\, S.\, Roessle\, M.\, White\, T. A.\, C hapman\, H. N.\, Meents\, A. & Oberthuer\, D. (2017). IUCrJ\n4\, 769.\n[3] Henkel\, A.\, Maracke\, J.\, Munke\, A.\, Galchenkova\, M.\, Rahmani Mash hour\, A.\, Reinke\, P.\, Do-\nmaracky\, M.\, Fleckenstein\, H.\, Hakanpä ä\, J.\, Meyer\, J.\, Tolstikova\, A.\, Carnis\, J.\, Middendorf\, P.\,\n Gelisio\, L.\, Yefanov\, O.\, Chapman\, H. N. & Oberthür\, D. (2022). Act a Cryst. A78\, e560.\n[4] White\, T. A.\, Mariani\, V.\, Brehm\, W.\, Yefa nov\, O.\, Barty\, A.\, Beyerlein\, K. R.\, Chervinskii\, F.\,\nGalli\, L. \, Gati\, C.\, Nakane\, T.\, Tolstikova\, A.\, Yamashita\, K.\, Yoon\, C. H.\, Diederichs\, K. & Chapman\,\nH. N. (2016). J. Appl. Cryst. 49\, 680.\ n[5] White\, T. A. Schoof\, T.\, Yakubov\, S.\, Tolstikova\, A.\, Middendo rf\, P.\, Karnevskiy\, M.\, Mariani\, V.\,\nHenkel\, A.\, Klopprogge\, B.\ , Hannappel\, J.\, Oberthür\, D.\, De Gennaro Aquino\, I.\, Egorov\, D.\, Munke\,\nA.\, Sprenger\, J.\, Pompidor\, G.\, Taberman\, H.\, Gruzinov\, A.\, Meyer\, J.\, Hakanpää\, J.\, & Gasthuber\,\nG. (2025). IUCrJ 12\, 9 7.\n[6] Mariani\, V.\, Morgan\, A.\, Yoon\, C. H.\, Lane\, T. J.\, White\, T. A.\, O’Grady\, C.\, Kuhn\, M.\, Aplin\, S.\,\nKoglin\, J.\, Barty\, A. & Chapman\, H. N. (2016). J. Appl. Cryst. 49\, 1073.\n\n\nCo-authors:\n Alessandra Henkel\, Julia Maracke\, Spyridon Chatziefthymiou\, Alexander G rebentsov\, Andrey Gruzinov\, Olga Merkulova\, Philipp Middendorf\, Alexan dra Tolstikova\, Thomas White\, Dominik Oberthür\, Johanna Hakanpää\n\n https://lindico453.srv.lu.se/event/583/contributions/1829/ LOCATION:LINXS at The Loop URL:https://lindico453.srv.lu.se/event/583/contributions/1829/ END:VEVENT BEGIN:VEVENT SUMMARY:Using advanced crystallographic approaches to resolve how orange c arotenoid protein photocycle works DTSTART;VALUE=DATE-TIME:20250923T073000Z DTEND;VALUE=DATE-TIME:20250923T080000Z DTSTAMP;VALUE=DATE-TIME:20260524T212503Z UID:indico-contribution-583-1846@lindico453.srv.lu.se DESCRIPTION:Speakers: Volha Chukhutsina (Vrije Universiteit Amsterdam)\nht tps://lindico453.srv.lu.se/event/583/contributions/1846/ LOCATION:LINXS at The Loop URL:https://lindico453.srv.lu.se/event/583/contributions/1846/ END:VEVENT BEGIN:VEVENT SUMMARY:Scientific opportunities for Serial Crystallography at ALBA synchr otron DTSTART;VALUE=DATE-TIME:20250922T162000Z DTEND;VALUE=DATE-TIME:20250922T163000Z DTSTAMP;VALUE=DATE-TIME:20260524T212503Z UID:indico-contribution-583-1843@lindico453.srv.lu.se DESCRIPTION:Speakers: Isidro Crespo (ALBA-CELLS)\, Xavi Carpena i Vilella (ALBA-CELLS)\nThe ALBA Synchrotron\, the 3 GeV light source in Southern Eu rope\, is preparing for its upgrade to a 4th generation storage ring (ALBA II) together with the construction of 3 beamlines and major upgrades on t he existing ones\, early next decade. Within this framework\, serial and t ime-resolved macromolecular crystallography (SSX and TR-MX) are identified as strategic growth areas.\nBL13-XALOC\, in operation since 2012 [1]\, ha s been the workhorse MX beamline at ALBA\, delivering photon fluxes up to 2.5 × 10¹² ph/s with beam sizes from 50 × 7 μm² to 300 × 100 μm² over 5.2–22 keV. The beamline has undergone an upgrade process including a Pilatus-3X 6M (100Hz) detector and a new automated sample changer. Besi des\, the beamline is equipped with a high viscosity extrusion injector fo r SSX experiments. The transition from single crystal oscillation MX to SS X is highly simplified thanks a three-axis motorized stage and sample extr usion is facilitated thanks to a PID pressure control system. Proof-of-con cept SSX experiments with test proteins have demonstrated compatibility wi th room-temperature data collection and negligible radiation damage [2]. P ump–probe TR-MX experiments using visible-light activation have also bee n successfully performed\, establishing a baseline for further development s [3].\nA second MX beamline\, BL06-XAIRA\, started regular user operation in June 2025\, offering a highly stable microfocus beam of 3 × 1 μm² a nd photon energy range of <4.0–14 keV. Equipped with a <60nm runout diff ractometer\, an EIGER2 XE 9M and a dual Channel-Cut/multilayer monochromat or [4]\, it enables high-flux\, high-stability microcrystallography. This equipment\, combined with the implementation in the following months of a fast (up to 750 mm/s) SSX stage\, designed for chips up to 60x40 mm will e nable fixed-target SSX experiments at higher time resolution. Background r eduction and long wavelength experiments are possible due to the recircula ted He environment enclosing the entire end station.\nThe ALBA II upgrade plan foresees substantial investment in SSX/TR-MX capabilities for both be amlines. For XALOC\, this includes hybrid tape-drive systems for tunable s oaking times\, and microfluidic injectors. XAIRA will focus on microfluidi c chips\, acoustic droplet ejection\, and conveyor-belt systems for contin uous fresh sample delivery. A pump-probe set up will be developed for both beamlines to allow for μs–ms TR-MX\, including choppers\, tunable lase rs\, and synchronization systems.\n[1] Juanhuix\, J.\, Gil-Ortiz\, F.\, Cu ní\, G.\, Colldelram\, C.\, Nicolas\, J.\, Lidon\, J.\, Boter\, E.\, Ruge t\, C.\, Ferrer S. & Benach\, J. (2014) Synchrotron Radiat. 21\, 679-689.\ n[2] Martin-Garcia J.M.\, Botha S.\, Hu H.\, Jernigan R.\, Castellví A.\, Lisova S.\, Gil-Ortiz F.\, Calisto B.\, Crespo I.\, Roy-Chowdhury S.\, Gr ieco A.\, Ketawala G.\, Weierstall U.\, Spence J.\, Fromme P.\, Zatsepin N .\, Boer D.R. & Carpena X. (2022) J. Synchrotron Radiat. 29(3): 896-\n[3] Kovalev K\, et al\, Nat Commun. 2020 May 1\;11(1):2137.\n[4] N. González et al.\, Proc. 12th Int. Conf. Mech. Eng. Design Synchrotron Radiat. Equip . Instrum. (MEDSI'23)\, Beijing\, China\, Nov. 2023\, pp.~5-9. doi:10.1842 9/JACoW-MEDSI2023-TUOAM04\n\n\nAuthors:\nXAVI CARPENA I VILELLA\, ISIDRO C RESPO\nCo-authors:\nDAMIÀ GARRIGA\, FERNANDO GIL-ORTIZ\, NAHIKARI GONZÁL EZ\, ALEIX TARRÉS\, ALBERT MIRET\, BERNAT MOLAS\, RICARDO VALCÁRCEL\, AL EJANDRO ENRIQUE\, CARLES COLLDELRAM\, IGORS ŠICS\, JOSÉ MARÍA ÁLVAREZ\ , MARCOS QUISPE\, JULIÁN ALBERTO GARCÍA\, ROELAND BOER\, JUDITH JUANHUIX \n\nhttps://lindico453.srv.lu.se/event/583/contributions/1843/ LOCATION:LINXS at The Loop URL:https://lindico453.srv.lu.se/event/583/contributions/1843/ END:VEVENT BEGIN:VEVENT SUMMARY:Current and future capabilities for serial and time resolved cryst allography at Diamond microfocus beamline VMXi DTSTART;VALUE=DATE-TIME:20250922T150000Z DTEND;VALUE=DATE-TIME:20250922T151500Z DTSTAMP;VALUE=DATE-TIME:20260524T212503Z UID:indico-contribution-583-1832@lindico453.srv.lu.se DESCRIPTION:Speakers: Juan Sanchez Weatherby (Diamond Light Source)\nThere is increasing interest in obtaining room temperature\, time resolved crys tal structures of\nproteins carrying out their biological functions. The t ransition between conventional cryogenic\nmacromolecular crystallography a nd serial crystallography involving microcrystals remains chal-\nlenging f or many projects. We have recently demonstrated the capability to measure good quality\,\nlow dose serial crystallography data from microcrystals wi thin crystallisation plates [1]. This capa-\nbility is available in the st andard operation mode of the beamline and does not require any specific\ns erial crystallography apparatus. In this approach\, microcrystals are tran sferred into a crystallisa-\ntion plate (typically 100 nL per drop) and ea ch droplet is subjected to raster scanning with a still\ndiffraction image measured every 10 µm. The resulting images are processed using standard serial\ndata processing software such as xia2.multiplex. This approach ena bles straightforward structure\ndetermination and analysis of crystal qual ity and unit cell parameters from non-optimised crys-\ntallisation conditi ons\, guiding users in their optimisation efforts. Very small quantities o f protein\nare required\, and the determination of a human peroxidase stru cture to 1.88 Angstrom resolution\nusing only 1.2 µL microcrystal suspens ion.\nSeveral approaches to sample delivery for time resolved crystallogra phy have been developed in-\ncluding droplet-on-demand tape drive-based sy stems developed for XFEL experiments that have\nbeen combined with X-ray e mission spectroscopy (XES) to monitor the redox and spin state of\nmetal - containing cofactors within the proteins [2]. However\, currently availabl e systems require\na large quantity of microcrystal sample as well as requ iring multiple skilled staff to operate. A\nnew system for serial crystall ography at VMXi is currently under development. This incorporates\na picol itre droplet-on-demand tape drive system capable of anaerobic operation to gether with an\nXES von Hamos spectrometer to enable spectroscopic validat ion in time resolved experiments of\nmetalloproteins. A compact design was required due to the tight spatial constraints of the VMXi\nend station th at was built for highly automated data collection from crystallisation pla tes\, and the\ndesign incorporates automation to reduce the number of pers onnel required to more closely ap-\nproach a typical synchrotron experimen t.\nProof of concept data obtained during the development process of the t ape drive and XES spec-\ntrometer will be presented\, including a high-res olution protein structure determined using the tape\ndrive system and XES data obtained from microcrystals of the copper enzyme nitrite reductase.\n [1] A.J. Thompson\, J. Sanchez-Weatherby\, L.J. Williams\, H. Mikolajek\, J. Sandy\, J.A.R. Worrall and\nM.A. Hough (2024) Efficient in situ screeni ng of and data collection from microcrystals in crystal-\nlization plates Acta Cryst.D80\, 279-288\n[2] Butyrin\, A. et al (2021) An on-demand\, dro p-on-drop method for studying enzyme catalysis by\nserial crystallography. Nature Methods 12\, 4461.\n\n\nAuthors:\nJuan Sanchez Weatherby\, Pierre Aller\, Amy Thompson\, Abby Telfer\, John Sutter\, James Sandy\, Halina Mi kolajek\, Matthew Rodrigues\, Mike Hough\, Allen Orville\n\nhttps://lindic o453.srv.lu.se/event/583/contributions/1832/ LOCATION:LINXS at The Loop URL:https://lindico453.srv.lu.se/event/583/contributions/1832/ END:VEVENT BEGIN:VEVENT SUMMARY:Welcome (Science Director Joachim Schnadt & MicroMAX Spokesperson Richard Neutze) DTSTART;VALUE=DATE-TIME:20250922T110000Z DTEND;VALUE=DATE-TIME:20250922T111000Z DTSTAMP;VALUE=DATE-TIME:20260524T212503Z UID:indico-contribution-583-1823@lindico453.srv.lu.se DESCRIPTION:Speakers: Joachim Schnadt (MAX IV Laboratory)\, Richard Neutze (University of Gothenburg)\nhttps://lindico453.srv.lu.se/event/583/contri butions/1823/ LOCATION:LINXS at The Loop URL:https://lindico453.srv.lu.se/event/583/contributions/1823/ END:VEVENT BEGIN:VEVENT SUMMARY:Lunch DTSTART;VALUE=DATE-TIME:20250922T100000Z DTEND;VALUE=DATE-TIME:20250922T110000Z DTSTAMP;VALUE=DATE-TIME:20260524T212503Z UID:indico-contribution-583-1827@lindico453.srv.lu.se DESCRIPTION:https://lindico453.srv.lu.se/event/583/contributions/1827/ LOCATION:LINXS at The Loop URL:https://lindico453.srv.lu.se/event/583/contributions/1827/ END:VEVENT BEGIN:VEVENT SUMMARY:Registration DTSTART;VALUE=DATE-TIME:20250922T090000Z DTEND;VALUE=DATE-TIME:20250922T100000Z DTSTAMP;VALUE=DATE-TIME:20260524T212503Z UID:indico-contribution-583-1826@lindico453.srv.lu.se DESCRIPTION:https://lindico453.srv.lu.se/event/583/contributions/1826/ LOCATION:LINXS at The Loop URL:https://lindico453.srv.lu.se/event/583/contributions/1826/ END:VEVENT BEGIN:VEVENT SUMMARY:Some questions can only be answered by time: how time-resolved cry stallography reshapes structural biology DTSTART;VALUE=DATE-TIME:20250922T111000Z DTEND;VALUE=DATE-TIME:20250922T114000Z DTSTAMP;VALUE=DATE-TIME:20260524T212503Z UID:indico-contribution-583-1824@lindico453.srv.lu.se DESCRIPTION:Speakers: Tobias Weinert (Paul Scherrer Institute)\nCryo-cryst allography was the driving force of structural biology\, literally deciphe ring the structures of life. The integrated knowledge has informed powerfu l machine learning technologies to not only solve the folding problem but to imagine new proteins\, some of which are even functional. However\, pro tein function is often dominated by small kinetic barriers that are not ea sily predicted. Time-resolved crystallography has set out to answer new qu estions about protein function and to resolve atomic motion on the femtose cond scale and kinetic intermediates ranging from femtoseconds to seconds - hopefully culminating in the ability to design molecular machines one da y. The talk aims to embed some of our research into this greater context.\ n\nhttps://lindico453.srv.lu.se/event/583/contributions/1824/ LOCATION:LINXS at The Loop URL:https://lindico453.srv.lu.se/event/583/contributions/1824/ END:VEVENT END:VCALENDAR