Hiroki Makita

University of Saskatchewan

Abstract:

Title 
X-ray Emission Spectroscopy on Metalloenzymes at Room Temperature: Combined XES and XRD Studies 
 
Hiroki Makita1, Patrick Rabe2, Vivek Srinivas3, Rahul Banerjee4, Franklin Fuller5, Roberto Alonso-Mori5, 
Leland Gee5, Philipp Simon1, Isabel Bogacz1, Insik Kim1, Kyle D. Sutherlin1, Jason Jones4, Jos Kamps2, 
Agatha  Butryn6,  Pierre  Aller6,  Andrea  Schmidt7,  Michal  Szcepek7,  Asmit  Bhowmick1,  Stephen  Keable1, 
Thomas Fransson8, Margaret Doyle1, Aaron Brewster1, Nicholas Sauter1, Alex Batyuk5, Uwe Bergmann5,9, 
John  Lipscomb4,  Martin  Högbom3,  Christopher  Schofield2,  Patrick  Scheerer7,  Allen  Orville6,  Vittal 
Yachandra1, Junko Yano1, Jan Kern1 
 1Lawrence  Berkeley  National  Laboratory,  2University  of  Oxford,  3Stockholm  University,  4University  of 
Minnesota, 5SLAC National Accelerator Laboratory, 6Diamond Light Source, 7Charité–
Universitätsmedizin Berlin, 8Heidelberg University, 9University of Wisconsin–Madison 
 
Abstract 
To achieve a detailed mechanistic understanding, the metalloenzyme catalysis is ideally studied under the 
physiological  conditions,  in  real  time  as  the  reactions  turn  over.    While  x-ray  serves  as  a  direct  metal-
specific  probe  that  monitors  electronic  structure  of  the  metal  center,  application  for  the  studies  of 
metalloenzyme  at room temperature is  challenging due  to poor sensitivity and  radiation damage.  X-ray 
Free  Electron  Lasers  (XFELs)  provide  the  high  brilliance  and  the  ultrafast  pulses  required  to  overcome 
these difficulties, and make time-resolved x-ray studies of metalloenzymes at room temperature feasible. 
 
Here, we present recent results on a series of oxygen-sensitive metalloenzymes at room temperature studied 
using XFEL.  Drop-on-Tape system was deployed to efficiently deliver the sample in-sync to the XFEL 
pulse.  In addition to minimizing the sample consumption rate, this sample delivery method allows for the 
simultaneous collection of X-ray diffraction (XRD) and X-ray emission (XES) data on metalloenzyme at 
room temperature, with ability to provide in-situ photo/gas/liquid activation for time-resolved studies.  In 
this  poster,  insight  from  the  time-resolved  XES  studies  on  Fe  and  Ni  containing  metalloenzymes  are 
featured.  

Poster Session Link:

https://gather.town/invite?token=0pEoq7VP

If you have any questions for the presenter, please contact them by either one of the following ways:

Hiroki Makita Community Page Link

Email: hmakita@lbl.gov