PT  - JOURNAL ARTICLE
AU  - Jose Olmos
AU  - Suraj Pandey
AU  - Jose M. Martin-Garcia
AU  - George Calvey
AU  - Andrea Katz
AU  - Juraj Knoska
AU  - Christopher Kupitz
AU  - Mark S. Hunter
AU  - Mengning Liang
AU  - Dominik Oberthuer
AU  - Oleksandr Yefanov
AU  - Max Wiedorn
AU  - Michael Heyman
AU  - Mark Holl
AU  - Kanupriya Pande
AU  - Anton Barty
AU  - Mitchell D. Miller
AU  - Stephan Stern
AU  - Shatabdi Roy-Chowdhury
AU  - Jesse Coe
AU  - Nirupa Nagaratnam
AU  - James Zook
AU  - Jacob Verburgt
AU  - Tyler Norwood
AU  - Ishwor Poudyal
AU  - David Xu
AU  - Jason Koglin
AU  - Matt Seaberg
AU  - Yun Zhao
AU  - Saša Bajt
AU  - Thomas Grant
AU  - Valerio Mariani
AU  - Garrett Nelson
AU  - Ganesh Subramanian
AU  - Euiyoung Bae
AU  - Raimund Fromme
AU  - Russel Fung
AU  - Peter Schwander
AU  - Matthias Frank
AU  - Thomas White
AU  - Uwe Weierstall
AU  - Nadia Zatsepin
AU  - John Spence
AU  - Petra Fromme
AU  - Henry N. Chapman
AU  - Lois Pollack
AU  - Lee Tremblay
AU  - Abbas Ourmazd
AU  - George N. Phillips, Jr.
AU  - Marius Schmidt
TI  - Enzyme Intermediates Captured “on-the-fly” by Mix-and-Inject Serial Crystallography
AID  - 10.1101/202432
DP  - 2018 Jan 01
TA  - bioRxiv
PG  - 202432
4099  - http://biorxiv.org/content/early/2018/01/18/202432.short
4100  - http://biorxiv.org/content/early/2018/01/18/202432.full
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.