3K0O

Room temperature structure of CypA mutant Ser99Thr

3K0O の概要

• エントリーDOI: 10.2210/pdb3k0o/pdb
• 関連するPDBエントリー: 3K0M 3K0N 3K0P 3K0Q 3K0R
• 分子名称: Cyclophilin A (2 entities in total)
• 機能のキーワード: proline isomerase, cyclosporin, host-virus interaction, isomerase, isopeptide bond, phosphoprotein, rotamase
• 由来する生物種: Homo sapiens (human)
• 細胞内の位置: Cytoplasm: P62937
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 18050.53

構造登録者

Fraser, J.S.,Alber, T. (登録日: 2009-09-24, 公開日: 2009-12-08, 最終更新日: 2023-09-06)

主引用文献

Fraser, J.S.,Clarkson, M.W.,Degnan, S.C.,Erion, R.,Kern, D.,Alber, T.
Hidden alternative structures of proline isomerase essential for catalysis.
Nature, 462:669-673, 2009

PubMed Abstract: A long-standing challenge is to understand at the atomic level how protein dynamics contribute to enzyme catalysis. X-ray crystallography can provide snapshots of conformational substates sampled during enzymatic reactions, while NMR relaxation methods reveal the rates of interconversion between substates and the corresponding relative populations. However, these current methods cannot simultaneously reveal the detailed atomic structures of the rare states and rationalize the finding that intrinsic motions in the free enzyme occur on a timescale similar to the catalytic turnover rate. Here we introduce dual strategies of ambient-temperature X-ray crystallographic data collection and automated electron-density sampling to structurally unravel interconverting substates of the human proline isomerase, cyclophilin A (CYPA, also known as PPIA). A conservative mutation outside the active site was designed to stabilize features of the previously hidden minor conformation. This mutation not only inverts the equilibrium between the substates, but also causes large, parallel reductions in the conformational interconversion rates and the catalytic rate. These studies introduce crystallographic approaches to define functional minor protein conformations and, in combination with NMR analysis of the enzyme dynamics in solution, show how collective motions directly contribute to the catalytic power of an enzyme.

PubMed: 19956261
DOI: 10.1038/nature08615

実験手法

X-RAY DIFFRACTION (1.55 Å)