8VQ1

Pseudomonas fluorescens G150T isocyanide hydratase at 298 K XFEL data, thioimidate intermediate

「8TSN」から置き換えられました

8VQ1 の概要

• エントリーDOI: 10.2210/pdb8vq1/pdb
• 関連するPDBエントリー: 8TSN 8TSU 8TSX 8TSY 8TSZ 8TT0 8TT1 8TT2 8TT4 8TT5 8VPW
• 分子名称: Isonitrile hydratase InhA, N-(4-nitrophenyl)methanimine, CHLORIDE ION, ... (4 entities in total)
• 機能のキーワード: isocyanide, isonitrile, x-ray free electron laser, serial crystallography, lyase
• 由来する生物種: Pseudomonas fluorescens
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 24410.29

構造登録者

Wilson, M.A.,Smith, N.,Dasgupta, M.,Dolamore, C. (登録日: 2024-01-17, 公開日: 2024-02-21, 最終更新日: 2024-10-09)

主引用文献

Smith, N.,Dasgupta, M.,Wych, D.C.,Dolamore, C.,Sierra, R.G.,Lisova, S.,Marchany-Rivera, D.,Cohen, A.E.,Boutet, S.,Hunter, M.S.,Kupitz, C.,Poitevin, F.,Moss 3rd, F.R.,Mittan-Moreau, D.W.,Brewster, A.S.,Sauter, N.K.,Young, I.D.,Wolff, A.M.,Tiwari, V.K.,Kumar, N.,Berkowitz, D.B.,Hadt, R.G.,Thompson, M.C.,Follmer, A.H.,Wall, M.E.,Wilson, M.A.
Changes in an enzyme ensemble during catalysis observed by high-resolution XFEL crystallography.
Sci Adv, 10:eadk7201-eadk7201, 2024

PubMed Abstract: Enzymes populate ensembles of structures necessary for catalysis that are difficult to experimentally characterize. We use time-resolved mix-and-inject serial crystallography at an x-ray free electron laser to observe catalysis in a designed mutant isocyanide hydratase (ICH) enzyme that enhances sampling of important minor conformations. The active site exists in a mixture of conformations, and formation of the thioimidate intermediate selects for catalytically competent substates. The influence of cysteine ionization on the ICH ensemble is validated by determining structures of the enzyme at multiple pH values. Large molecular dynamics simulations in crystallo and time-resolved electron density maps show that Asp ionizes during catalysis and causes conformational changes that propagate across the dimer, permitting water to enter the active site for intermediate hydrolysis. ICH exhibits a tight coupling between ionization of active site residues and catalysis-activated protein motions, exemplifying a mechanism of electrostatic control of enzyme dynamics.

PubMed: 38536910
DOI: 10.1126/sciadv.adk7201

実験手法

X-RAY DIFFRACTION (1.3 Å)