SMBp Special Seminar: Radoslav Enchev (The Francis Crick Institute)
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America/New_York
3rd Floor Classroom (162 Fifth Avenue )
3rd Floor Classroom
162 Fifth Avenue
Description
Speaker: Radoslav Enchev
Title: Advancing the Development & Applications of Time-Resolved Cryo-EM
Abstract: Comprehensive understanding of the structural transitions that govern biochemical pathways is pivotal to fundamental and translational research. While cryo-EM and single-particle analysis theoretically offer access to all conformational states of the studied macromolecules, high-energy intermediates, which are of critical importance to the molecular mechanism, are substantially less likely to be observed under the steady state condition under which conventional sample preparation is performed. We have been developing an alternative to experimentally enrich such high energy reaction intermediates through time-resolved sample preparation. In particular, our lab has focused on developing and applying a method to observe biochemical processes with atomic spatial and millisecond time-resolution using cryo-EM. Biological macromolecules and/or chemical compounds are mixed and incubated in a microfluidics device on time scales relevant to binding and catalytic events. Droplets of these solutions are deposited onto a cryo-EM sample grid, which is flash-frozen into liquid ethane. Iterating the procedure at increasing incubation times after sample mixing, alongside single particle analysis, allows the visualization of a biochemical binding and/or enzymatic reaction as a time-lapse “movie”. I will present our current and future methods development efforts as well as applications to studying several biological systems.
Affiliation: R. I. Enchev, The Visual Biochemistry Laboratory, The Francis Crick Institute, London, UK
