RT Journal Article
SR Electronic
T1 Enzyme Intermediates Captured “on-the-fly” by Mix-and-Inject Serial Crystallography
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 202432
DO 10.1101/202432
A1 Jose Olmos
A1 Suraj Pandey
A1 Jose M. Martin-Garcia
A1 George Calvey
A1 Andrea Katz
A1 Juraj Knoska
A1 Christopher Kupitz
A1 Mark S. Hunter
A1 Mengning Liang
A1 Dominik Oberthuer
A1 Oleksandr Yefanov
A1 Max Wiedorn
A1 Michael Heyman
A1 Mark Holl
A1 Kanupriya Pande
A1 Anton Barty
A1 Mitchell D. Miller
A1 Stephan Stern
A1 Shatabdi Roy-Chowdhury
A1 Jesse Coe
A1 Nirupa Nagaratnam
A1 James Zook
A1 Jacob Verburgt
A1 Tyler Norwood
A1 Ishwor Poudyal
A1 David Xu
A1 Jason Koglin
A1 Matt Seaberg
A1 Yun Zhao
A1 Saša Bajt
A1 Thomas Grant
A1 Valerio Mariani
A1 Garrett Nelson
A1 Ganesh Subramanian
A1 Euiyoung Bae
A1 Raimund Fromme
A1 Russel Fung
A1 Peter Schwander
A1 Matthias Frank
A1 Thomas White
A1 Uwe Weierstall
A1 Nadia Zatsepin
A1 John Spence
A1 Petra Fromme
A1 Henry N. Chapman
A1 Lois Pollack
A1 Lee Tremblay
A1 Abbas Ourmazd
A1 George N. Phillips, Jr.
A1 Marius Schmidt
YR 2018
UL http://biorxiv.org/content/early/2018/01/18/202432.abstract
AB Ever since the first atomic structure of an enzyme was solved, the discovery of the mechanism and dynamics of reactions catalyzed by biomolecules has been the key goal for the understanding of the molecular processes that drive life on earth. Despite a large number of successful methods for trapping reaction intermediates, the direct observation of an ongoing reaction has been possible only in rare and exceptional cases. Here, we demonstrate a general method for capturing enzyme catalysis ‘in-action’ by ‘mix-and-inject serial crystallography’. Specifically, we follow the catalytic reaction of the Mycobacterium tuberculosis α-lactamase with the 3rd generation antibiotic ceftriaxone by time-resolved serial femtosecond crystallography. The results reveal, in near atomic detail, antibiotic cleavage and inactivation on the millisecond to second time scales including the crossover from transition state kinetics to steady-state kinetics.Synopsis An enzymatically catalyzed reaction is initiated by diffusion based mixing of substrate and followed at runtime by time-resolved serial crystallography using a free electron laser.