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Title: Structural enzymology using X-ray free electron lasers

Journal Article · · Structural Dynamics
DOI:https://doi.org/10.1063/1.4972069· OSTI ID:1395301
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  1. Univ. of Wisconsin, Milwaukee, WI (United States). Dept. of Physics
  2. Rice Univ., Houston, TX (United States). Dept. of BioSciences
  3. Arizona State Univ., Tempe, AZ (United States). Dept. of Physics
  4. Marbles Inc., Westfield, IN (United States)
  5. Deutsches Elektronen-Synchrotron (DESY), Hamburg (Germany). Center for Free-Electron Laser Science; Univ. of Hamburg (Germany)
  6. SLAC National Accelerator Lab., Menlo Park, CA (United States). Linac Coherent Light Source (LCLS)
  7. Cornell Univ., Ithaca, NY (United States). Dept. of Applied and Engineering Physics
  8. Deutsches Elektronen-Synchrotron (DESY), Hamburg (Germany). Center for Free-Electron Laser Science
  9. Univ. of Hamburg (Germany)
  10. State Univ. of New York (SUNY), Buffalo, NY (United States). Hauptman-Woodward Inst.
  11. Arizona State Univ., Tempe, AZ (United States). School of Molecular Sciences and Biodesign Center for Applied Structural Discovery
  12. 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
Citation Metrics:
Cited by: 74 works
Citation information provided by
Web of Science

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Cited By (35)

Non-Cryogenic Structure and Dynamics of HIV-1 Integrase Catalytic Core Domain by X-ray Free-Electron Lasers journal April 2019
Enzyme intermediates captured “on the fly” by mix-and-inject serial crystallography journal May 2018
High-viscosity injector-based pink-beam serial crystallography of microcrystals at a synchrotron radiation source text January 2019
Short-wavelength free-electron laser sources and science: a review journal October 2017
Snapshot of an oxygen intermediate in the catalytic reaction of cytochrome c oxidase journal February 2019
Outrunning damage: Electrons vs X-rays—timescales and mechanisms journal July 2017
Perspective: Opportunities for ultrafast science at SwissFEL text January 2017
Time-resolved serial femtosecond crystallography at the European XFEL text January 2019
Emission and dose characterization of the 1 kHz repetition rate high-Z metal K $$_{\alpha }$$ α source driven by 20 mJ femtosecond pulses journal February 2019
Microfluidics: From crystallization to serial time-resolved crystallography journal May 2017
Sample delivery for serial crystallography at free-electron lasers and synchrotrons journal January 2019
Perspective: Opportunities for ultrafast science at SwissFEL text January 2017
Capturing an initial intermediate during the P450nor enzymatic reaction using time-resolved XFEL crystallography and caged-substrate journal November 2017
Perspective: Opportunities for ultrafast science at SwissFEL journal November 2017
On-chip crystallization for serial crystallography experiments and on-chip ligand-binding studies text January 2019
On-chip crystallization for serial crystallography experiments and on-chip ligand-binding studies text January 2019
Rapid sample delivery for megahertz serial crystallography at X-ray FELs text January 2018
Structure of the 30S ribosomal decoding complex at ambient temperature journal August 2018
High-viscosity injector-based pink-beam serial crystallography of microcrystals at a synchrotron radiation source journal April 2019
Membrane protein megahertz crystallography at the European XFEL text January 2019
Shining light on cysteine modification: connecting protein conformational dynamics to catalysis and regulation journal June 2019
Time-resolved x-ray crystallography capture of a slow reaction tetrahydrofolate intermediate journal March 2019
Membrane protein megahertz crystallography at the European XFEL journal November 2019
Watching proteins function with time-resolved x-ray crystallography journal August 2017
Pink-beam serial crystallography journal November 2017
Ultracompact 3D microfluidics for time-resolved structural biology text January 2020
Rapid Sample Delivery for Megahertz Serial Crystallography at X-ray FELs text January 2018
Short-wavelength free-electron laser sources and science: a review text January 2017
Ultracompact 3D microfluidics for time-resolved structural biology journal January 2020
Molecular dynamics simulations of macromolecular crystals journal November 2018
Mix-and-inject XFEL crystallography reveals gated conformational dynamics during enzyme catalysis journal December 2019
Time-resolved serial femtosecond crystallography at the European XFEL journal November 2019
Membrane protein megahertz crystallography at the European XFEL text January 2019
Rapid sample delivery for megahertz serial crystallography at X-ray FELs journal July 2018
Ultracompact 3D microfluidics for time-resolved structural biology text January 2020

Figures / Tables (2)