7RY4

Multi-conformer model of Ketosteroid Isomerase Y57F/D40N mutant from Pseudomonas Putida (pKSI) bound to a transition state analog at 250 K

7RY4 の概要

• エントリーDOI: 10.2210/pdb7ry4/pdb
• 関連するPDBエントリー: 7RXF 7RXK
• 分子名称: Steroid Delta-isomerase, CHLORIDE ION, MAGNESIUM ION, ... (5 entities in total)
• 機能のキーワード: isomerase
• 由来する生物種: Pseudomonas putida (Arthrobacter siderocapsulatus)
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 29820.50

構造登録者

Yabukarski, F.,Doukov, T.,Herschlag, D. (登録日: 2021-08-24, 公開日: 2022-11-09, 最終更新日: 2023-11-22)

主引用文献

Yabukarski, F.,Doukov, T.,Pinney, M.M.,Biel, J.T.,Fraser, J.S.,Herschlag, D.
Ensemble-function relationships to dissect mechanisms of enzyme catalysis.
Sci Adv, 8:eabn7738-eabn7738, 2022

PubMed Abstract: Decades of structure-function studies have established our current extensive understanding of enzymes. However, traditional structural models are snapshots of broader conformational ensembles of interchanging states. We demonstrate the need for conformational ensembles to understand function, using the enzyme ketosteroid isomerase (KSI) as an example. Comparison of prior KSI cryogenic x-ray structures suggested deleterious mutational effects from a misaligned oxyanion hole catalytic residue. However, ensemble information from room-temperature x-ray crystallography, combined with functional studies, excluded this model. Ensemble-function analyses can deconvolute effects from altering the probability of occupying a state (-effects) and changing the reactivity of each state (-effects); our ensemble-function analyses revealed functional effects arising from weakened oxyanion hole hydrogen bonding and substrate repositioning within the active site. Ensemble-function studies will have an integral role in understanding enzymes and in meeting the future goals of a predictive understanding of enzyme catalysis and engineering new enzymes.

PubMed: 36240280
DOI: 10.1126/sciadv.abn7738

実験手法

X-RAY DIFFRACTION (1.11 Å)