5HFO

CRYSTAL STRUCTURE OF OXA-232 BETA-LACTAMASE

5HFO の概要

• エントリーDOI: 10.2210/pdb5hfo/pdb
• 分子名称: Beta-lactamase, SULFATE ION, GLYCEROL, ... (5 entities in total)
• 機能のキーワード: class d beta-lactamase oxa-232, antibiotic, dimer, hydrolase
• 由来する生物種: Escherichia coli
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 58150.81

構造登録者

Retailleau, P.,Oueslati, S.,Cisse, C.,Nordmann, P.,Naas, T.,Iorga, B. (登録日: 2016-01-07, 公開日: 2017-01-18, 最終更新日: 2024-01-10)

主引用文献

Oueslati, S.,Retailleau, P.,Marchini, L.,Berthault, C.,Dortet, L.,Bonnin, R.A.,Iorga, B.I.,Naas, T.
Role of Arginine 214 in the Substrate Specificity of OXA-48.
Antimicrob.Agents Chemother., 64:-, 2020

PubMed Abstract: Increasing numbers of variants of the carbapenem-hydrolyzing class D β-lactamase OXA-48 are identified in worldwide. Among them, OXA-181 and OXA-232 are of particular interest, as they differ from each other by a single amino acid substitution at position 214 (R in OXA-181 and S in OXA-232) that results in reduced carbapenem-hydrolyzing activity for OXA-232. To investigate the role of amino acid position 214 (AA214), the X-ray structure of OXA-232 was determined and AA214 of OXA-48 and of OXA-232 was replaced by G, L, D, E, S, R, and K using site-directed mutagenesis. These mutants were phenotypically characterized, and three mutants of OXA-232 were purified to study their steady-state kinetic properties. The X-ray structure of OXA-232 along with molecular modeling studies showed that the interaction via a salt bridge between R214 and D159 in OXA-48 is not possible with the G214 or S214 mutation. In contrast, with K214, which is also positively charged, the interaction with D159 is maintained. With the E214 mutant, an alternative binding conformation of imipenem that is not compatible with a nucleophilic attack by S70 was evidenced. Thus, imipenem has a very poor apparent affinity for the E214 mutant because of its nonproductive binding mode. Similarly, we could explain the lack of temocillin hydrolysis by the OXA-232-S214E mutant, which is due to the unfavorable interaction between the negatively charged R1 substituent of temocillin with the E214 residue. Overall, we demonstrate that AA214 in OXA-48-like β-lactamases is critical for the carbapenemase activity.

PubMed: 32071047
DOI: 10.1128/AAC.02329-19

実験手法

X-RAY DIFFRACTION (2.21 Å)