Structural enzymology using X-ray free electron lasers
- Univ. of Wisconsin, Milwaukee, WI (United States). Dept. of Physics
- Rice Univ., Houston, TX (United States). Dept. of BioSciences
- Arizona State Univ., Tempe, AZ (United States). Dept. of Physics
- Marbles Inc., Westfield, IN (United States)
- Deutsches Elektronen-Synchrotron (DESY), Hamburg (Germany). Center for Free-Electron Laser Science; Univ. of Hamburg (Germany)
- SLAC National Accelerator Lab., Menlo Park, CA (United States). Linac Coherent Light Source (LCLS)
- Cornell Univ., Ithaca, NY (United States). Dept. of Applied and Engineering Physics
- Deutsches Elektronen-Synchrotron (DESY), Hamburg (Germany). Center for Free-Electron Laser Science
- Univ. of Hamburg (Germany)
- State Univ. of New York (SUNY), Buffalo, NY (United States). Hauptman-Woodward Inst.
- Arizona State Univ., Tempe, AZ (United States). School of Molecular Sciences and Biodesign Center for Applied Structural Discovery
- Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States)
Mix-and-inject serial crystallography (MISC) is a technique designed to image enzyme catalyzed reactions in which small protein crystals are mixed with a substrate just prior to being probed by an X-ray pulse. This approach offers several advantages over flow cell studies. It provides (i) room temperature structures at near atomic resolution, (ii) time resolution ranging from microseconds to seconds, and (iii) convenient reaction initiation. It outruns radiation damage by using femtosecond X-ray pulses allowing damage and chemistry to be separated. Here, we demonstrate that MISC is feasible at an X-ray free electron laser by studying the reaction of M. tuberculosis ß-lactamase microcrystals with ceftriaxone antibiotic solution. Electron density maps of the apo-ß-lactamase and of the ceftriaxone bound form were obtained at 2.8 Å and 2.4 Å resolution, respectively. Finally, these results pave the way to study cyclic and non-cyclic reactions and represent a new field of time-resolved structural dynamics for numerous substrate-triggered biological reactions.
- Research Organization:
- Lawrence Livermore National Laboratory (LLNL), Livermore, CA (United States); SLAC National Accelerator Laboratory (SLAC), Menlo Park, CA (United States)
- Sponsoring Organization:
- USDOE Office of Science (SC), Basic Energy Sciences (BES); National Institutes of Health (NIH); National Science Foundation (NSF)
- Grant/Contract Number:
- AC52-07NA27344; AC02-76SF00515
- OSTI ID:
- 1395301
- Alternate ID(s):
- OSTI ID: 1369303; OSTI ID: 2229580
- Report Number(s):
- LLNL-JRNL-857682; SDTYAE
- Journal Information:
- Structural Dynamics, Vol. 4, Issue 4; ISSN 2329-7778
- Publisher:
- American Crystallographic Association/AIPCopyright Statement
- Country of Publication:
- United States
- Language:
- English
Web of Science
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