Journal:IUCrJ:S205225252000072X

From Proteopedia

(Difference between revisions)
Jump to: navigation, search
Line 9: Line 9:
Our model system for this work was Cyclophilin A (CypA), which we had previously studied at multiple temperatures. As it turned out, both microED and XFELs enabled us to solve high-resolution structures of CypA, but the conformations of the protein were not the same. In the microED structure, CypA adopted a single conformation previously seen in cryogenic X-ray studies, whereas the XFEL data revealed multiple conformations, as seen in previous room-temperature X-ray studies. So, our work reveals that temperature is a defining difference between structures solved using XFELs and those solved using microED. For many proteins, different conformations are visible at cryogenic temperatures when compared to room temperature. This means that these two methods provide complementary views into the molecular landscape, in addition to enabling collection of high-resolution data from small crystals.
Our model system for this work was Cyclophilin A (CypA), which we had previously studied at multiple temperatures. As it turned out, both microED and XFELs enabled us to solve high-resolution structures of CypA, but the conformations of the protein were not the same. In the microED structure, CypA adopted a single conformation previously seen in cryogenic X-ray studies, whereas the XFEL data revealed multiple conformations, as seen in previous room-temperature X-ray studies. So, our work reveals that temperature is a defining difference between structures solved using XFELs and those solved using microED. For many proteins, different conformations are visible at cryogenic temperatures when compared to room temperature. This means that these two methods provide complementary views into the molecular landscape, in addition to enabling collection of high-resolution data from small crystals.
 +
 +
Comparison of the 2mFoFc map and the refined multi-conformer model produced from each serial XFEL experiment. Maps were visualized at multiple contour levels to show evidence for alternative conformations. Following multi-conformer refinement, ensembles were generated from each model using phenix.ensemble_refine. In the right panel, a histogram of the Chi1 angles for residue 113 is plotted for the ensemble. Multi-conformer models plus maps, and the distribution of Chi1 angles across the ensemble models are similar for all three XFEL data sets:
 +
*<scene name='83/834718/Cv/2'>XFEL MESH</scene>.
<b>References</b><br>
<b>References</b><br>
<references/>
<references/>
</StructureSection>
</StructureSection>
__NOEDITSECTION__
__NOEDITSECTION__

Revision as of 14:56, 2 February 2020

Drag the structure with the mouse to rotate

Proteopedia Page Contributors and Editors (what is this?)

Alexander Berchansky, Jaime Prilusky

This page complements a publication in scientific journals and is one of the Proteopedia's Interactive 3D Complement pages. For aditional details please see I3DC.
Personal tools