BEGIN:VCALENDAR VERSION:2.0 PRODID:-//CERN//INDICO//EN BEGIN:VEVENT SUMMARY:Electrostatically Gated Enzyme Dynamics During Catalysis DTSTART;VALUE=DATE-TIME:20250922T143000Z DTEND;VALUE=DATE-TIME:20250922T150000Z DTSTAMP;VALUE=DATE-TIME:20260525T143038Z UID:indico-contribution-213-1831@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:20260525T143038Z UID:indico-contribution-213-1830@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:20260525T143038Z UID:indico-contribution-213-1828@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:20260525T143038Z UID:indico-contribution-213-1825@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:Serial crystallography at the beamline P11\, PETRA III DTSTART;VALUE=DATE-TIME:20250922T131000Z DTEND;VALUE=DATE-TIME:20250922T132500Z DTSTAMP;VALUE=DATE-TIME:20260525T143038Z UID:indico-contribution-213-1829@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: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:20260525T143038Z UID:indico-contribution-213-1832@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:20260525T143038Z UID:indico-contribution-213-1823@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: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:20260525T143038Z UID:indico-contribution-213-1824@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