Send to

Choose Destination
Struct Dyn. 2016 Dec 15;4(4):044003. doi: 10.1063/1.4972069. eCollection 2017 Jul.

Structural enzymology using X-ray free electron lasers.

Author information

Physics Department, University of Wisconsin-Milwaukee , 3135 N. Maryland Ave, Milwaukee, Wisconsin 53211, USA.
Department of BioSciences, Rice University , 6100 Main Street, Houston, Texas 77005, USA.
Department of Physics, Arizona State University , Tempe, Arizona 85287, USA.
Marbles Inc. , 1900 Belvedere Pl, Westfield, Indiana 46074, USA.
Linac Coherent Light Source, Stanford Linear Accelerator Center (SLAC) National Accelerator Laboratory , 2575 Sand Hill Road, Menlo Park, California 94025, USA.
Department of Applied and Engineering Physics, Cornell University , 254 Clark Hall, Ithaca, New York 14853, USA.
Center for Free-Electron Laser Science, DESY , Notkestrasse 85, 22607 Hamburg, Germany.
University of Hamburg , Luruper Chaussee 149, 22761 Hamburg, Germany.
Hauptman-Woodward Institute, State University of New York at Buffalo , 700 Ellicott Street, Buffalo, New York 14203, USA.
School of Molecular Sciences and Biodesign Center for Applied Structural Discovery, Arizona State University , Tempe, Arizona 85287-1604, USA.
Lawrence Livermore National Laboratory , Livermore, California 94550, USA.


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. 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.

Supplemental Content

Full text links

Icon for PubMed Central
Loading ...
Support Center