BEGIN:VCALENDAR VERSION:2.0 PRODID:-//CERN//INDICO//EN BEGIN:VEVENT SUMMARY:Wrap-up DTSTART;VALUE=DATE-TIME:20250924T094500Z DTEND;VALUE=DATE-TIME:20250924T100000Z DTSTAMP;VALUE=DATE-TIME:20260524T122247Z UID:indico-contribution-217-1861@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:20260524T122247Z UID:indico-contribution-217-1860@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:20260524T122247Z UID:indico-contribution-217-1859@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:20260524T122247Z UID:indico-contribution-217-1858@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:20260524T122247Z UID:indico-contribution-217-1857@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:20260524T122247Z UID:indico-contribution-217-1856@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 END:VCALENDAR