8ECF

Cryo structure of Mycobacterium tuberculosis beta lactamase microcrystals mixed with sulbactam for 3 hours

8ECF の概要

• エントリーDOI: 10.2210/pdb8ecf/pdb
• 分子名称: Beta-lactamase, PHOSPHATE ION, TRANS-ENAMINE INTERMEDIATE OF SULBACTAM, ... (4 entities in total)
• 機能のキーワード: beta lactamase, sulbactam, inhibitor, hydrolase, hydrolase-inhibitor complex, hydrolase/inhibitor
• 由来する生物種: Mycobacterium tuberculosis
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 114924.32

構造登録者

Malla, T.N.,Schmidt, M. (登録日: 2022-09-01, 公開日: 2023-03-08, 最終更新日: 2024-11-20)

主引用文献

Malla, T.N.,Zielinski, K.,Aldama, L.,Bajt, S.,Feliz, D.,Hayes, B.,Hunter, M.,Kupitz, C.,Lisova, S.,Knoska, J.,Martin-Garcia, J.M.,Mariani, V.,Pandey, S.,Poudyal, I.,Sierra, R.G.,Tolstikova, A.,Yefanov, O.,Yoon, C.H.,Ourmazd, A.,Fromme, P.,Schwander, P.,Barty, A.,Chapman, H.N.,Stojkovic, E.A.,Batyuk, A.,Boutet, S.,Phillips Jr., G.N.,Pollack, L.,Schmidt, M.
Heterogeneity in M. tuberculosis beta-lactamase inhibition by Sulbactam.
Nat Commun, 14:5507-5507, 2023

PubMed Abstract: For decades, researchers have elucidated essential enzymatic functions on the atomic length scale by tracing atomic positions in real-time. Our work builds on possibilities unleashed by mix-and-inject serial crystallography (MISC) at X-ray free electron laser facilities. In this approach, enzymatic reactions are triggered by mixing substrate or ligand solutions with enzyme microcrystals. Here, we report in atomic detail (between 2.2 and 2.7 Å resolution) by room-temperature, time-resolved crystallography with millisecond time-resolution (with timepoints between 3 ms and 700 ms) how the Mycobacterium tuberculosis enzyme BlaC is inhibited by sulbactam (SUB). Our results reveal ligand binding heterogeneity, ligand gating, cooperativity, induced fit, and conformational selection all from the same set of MISC data, detailing how SUB approaches the catalytic clefts and binds to the enzyme noncovalently before reacting to a trans-enamine. This was made possible in part by the application of singular value decomposition to the MISC data using a program that remains functional even if unit cell parameters change up to 3 Å during the reaction.

PubMed: 37679343
DOI: 10.1038/s41467-023-41246-1

実験手法

X-RAY DIFFRACTION (2.736 Å)