2024-03-29T09:08:55Zhttps://keep.lib.asu.edu/oai/requestoai:keep.lib.asu.edu:node-1303022021-08-16T19:23:30Zoai_pmh:all130302
https://hdl.handle.net/2286/R.I.44525
Kupitz, C., Olmos, J. L., Holl, M., Tremblay, L., Pande, K., Pandey, S., . . . Schmidt, M. (2017). Structural enzymology using X-ray free electron lasers. Structural Dynamics, 4(4), 044003. doi:10.1063/1.4972069
10.1063/1.4972069
2329-7778
http://rightsstatements.org/vocab/InC/1.0/
http://creativecommons.org/licenses/by/4.0
2016-12-15
7 pages
eng
Kupitz, Christopher
Olmos, Jose L.
Holl, Mark
Tremblay, Lee
Pande, Kanupriya
Pandey, Suraj
Oberthur, Dominik
Hunter, Mark
Liang, Mengning
Aquila, Andrew
Tenboer, Jason
Calvey, George
Katz, Andrea
Chen, Yujie
Wiedorn, Max O.
Knoska, Juraj
Meents, Alke
Majriani, Valerio
Norwood, Tyler
Poudyal, Ishwor
Grant, Thomas
Miller, Mitchell D.
Xu, Weijun
Tolstikova, Aleksandra
Morgan, Andrew
Metz, Markus
Martin Garcia, Jose Manuel
Zook, James
Roy Chowdhury, Shatabdi
Coe, Jesse
Nagaratnam, Nirupa
Meza-Aguilar, Domingo
Fromme, Raimund
Basu, Shibom
Frank, Matthias
White, Thomas
Barty, Anton
Bajt, Sasa
Yefanov, Oleksandr
Chapman, Henry N.
Zatsepin, Nadia
Nelson, Garrett
Weierstall, Uwe
Spence, John
Schwander, Peter
Pollack, Lois
Fromme, Petra
Ourmazd, Abbas
Phillips, George N.
Schmidt, Marius
College of Liberal Arts and Sciences
Department of Physics
School of Molecular Sciences
Biodesign Institute
Applied Structural Discovery
Text
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.
Structural enzymology using X-ray free electron lasers