TY - JOUR T1 - Mapping the Conformational Landscape of a Dynamic Enzyme by XFEL and Multitemperature Crystallography JF - bioRxiv DO - 10.1101/016733 SP - 016733 AU - Daniel A. Keedy AU - Lillian R. Kenner AU - Matthew Warkentin AU - Rahel A. Woldeyes AU - Michael C. Thompson AU - Aaron S. Brewster AU - Andrew H. Van Benschoten AU - Elizabeth L. Baxter AU - Jesse B. Hopkins AU - Monarin Uervirojnangkoorn AU - Scott E. McPhillps AU - Jinhu Song AU - Roberto Alonso-Mori AU - James M. Holton AU - William I. Weis AU - Axel T. Brunger AU - S. Michael Soltis AU - Henrik Lemke AU - Ana Gonzalez AU - Nicholas K. Sauter AU - Aina E. Cohen AU - Henry van den Bedem AU - Robert E. Thorne AU - James Fraser Y1 - 2015/01/01 UR - http://biorxiv.org/content/early/2015/03/19/016733.abstract N2 - 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. ER -