6KPH

343 K cryoEM structure of Sso-KARI in complex with Mg2+, NADH and CPD

6KPH の概要

• エントリーDOI: 10.2210/pdb6kph/pdb
• 関連するPDBエントリー: 6KOU
• EMDBエントリー: 0746
• 分子名称: Ketol-acid reductoisomerase, MAGNESIUM ION, 1,4-DIHYDRONICOTINAMIDE ADENINE DINUCLEOTIDE, ... (4 entities in total)
• 機能のキーワード: complex, isomerase
• 由来する生物種: Saccharolobus solfataricus
• タンパク質・核酸の鎖数: 12
• 化学式量合計: 456888.06

構造登録者

Chen, C.Y.,Chang, Y.C.,Lin, B.L.,Huang, C.H.,Tsai, M.D. (登録日: 2019-08-15, 公開日: 2020-03-25, 最終更新日: 2024-03-27)

主引用文献

Chen, C.Y.,Chang, Y.C.,Lin, B.L.,Huang, C.H.,Tsai, M.D.
Temperature-Resolved Cryo-EM Uncovers Structural Bases of Temperature-Dependent Enzyme Functions.
J.Am.Chem.Soc., 141:19983-19987, 2019

PubMed Abstract: Protein functions are temperature-dependent, but protein structures are usually solved at a single (often low) temperature because of limitations on the conditions of crystal growth or protein vitrification. Here we demonstrate the feasibility of solving cryo-EM structures of proteins vitrified at high temperatures, solve 12 structures of an archaeal ketol-acid reductoisomerase (KARI) vitrified at 4-70 °C, and show that structures of both the Mg form (KARI:2Mg) and its ternary complex (KARI:2Mg:NADH:inhibitor) are temperature-dependent in correlation with the temperature dependence of enzyme activity. Furthermore, structural analyses led to dissection of the induced-fit mechanism into ligand-induced and temperature-induced effects and to capture of temperature-resolved intermediates of the temperature-induced conformational change. The results also suggest that it is preferable to solve cryo-EM structures of protein complexes at functional temperatures. These studies should greatly expand the landscapes of protein structure-function relationships and enhance the mechanistic analysis of enzymatic functions.

PubMed: 31829582
DOI: 10.1021/jacs.9b10687

実験手法

ELECTRON MICROSCOPY (2.41 Å)