0 Citation(s)

0 questions (Ask a question)

Mapping the Conformational Landscape of a Dynamic Enzyme by XFEL and Multitemperature Crystallography

By Daniel A Keedy, Lillian R Kenner, Matthew Warkentin, Rahel Woldeyes1, Michael Thompson1, Aaron S Brewster, Andrew H Van Benschoten, Elizabeth L Baxter, Jesse B Hopkins, Monarin Uervirojnangkoorn, Scott E McPhillps, Jinhu Song, Roberto Alonso-Mori, James Holton2, William I Weis, Axel T Brunger, S. Michael Soltis, Henrik Lemke, Ana Gonzalez, Nicholas K Sauter, Aina E Cohen, Henry Van Den Bedem, Robert E Thorne, James Fraser3

1. University of California - San Francisco 2. University of California San Francisco 3. University of California-San Francisco

See also

No results found.

Category

BioXFEL Publications

Published on

Type

posted-content

Author

Daniel A Keedy and Lillian R Kenner and Matthew Warkentin and Rahel Woldeyes and Michael C Thompson and Aaron S Brewster and Andrew H Van Benschoten and Elizabeth L Baxter and Jesse B Hopkins and Monarin Uervirojnangkoorn and Scott E McPhillps and Jinhu Song and Roberto Alonso-Mori and James M Holton and William I Weis and Axel T Brunger and S. Michael Soltis and Henrik Lemke and Ana Gonzalez and Nicholas K Sauter and Aina E Cohen and Henry van den Bedem and Robert E Thorne and James Fraser

Citation

Keedy, D.A. et al., 2015. Mapping the Conformational Landscape of a Dynamic Enzyme by XFEL and Multitemperature Crystallography. Available at: http://dx.doi.org/10.1101/016733.

Abstract

Determining the interconverting conformations of dynamic proteins in atomic detail is a major challenge for structural biology. Conformational heterogeneity in the active site of the dynamic enzyme cyclophilin A (CypA) has been previously linked to its catalytic function. Here we compare the conformational ensembles of CypA by fixed-target X-ray free electron laser (XFEL) crystallography and multitemperature synchrotron crystallography. The “diffraction-before-destruction” nature of XFEL experiments provides a radiation-damage-free view of the functionally important alternative conformations of CypA. We monitored the temperature dependences of these alternative conformations with eight synchrotron datasets spanning 100-310 K. Multiconformer models show that many alternative conformations in CypA are populated above, but not below, the glass transition temperature (~200 K) and reveal abrupt changes in protein flexibility that provide all-atom insight into conformational coupling. Together, our XFEL data and multitemperature analyses motivate a new generation of time-resolved experiments to structurally characterize the dynamic underpinnings of protein function.  

DOI

http://dx.doi.org/10.1101/016733

Funding

NSF-STC Biology with X-ray Lasers (NSF-1231306)