5KSH

Crystal structure of penicillin-binding protein 2 from Neisseria gonorrhoeae containing an A501T mutation associated with cephalosporin resistance

5KSH の概要

• エントリーDOI: 10.2210/pdb5ksh/pdb
• 分子名称: Penicillin-binding protein 2, GLYCEROL, SULFATE ION, ... (4 entities in total)
• 機能のキーワード: biosynthetic protein, class b transpeptidase, peptidoglycan synthesis
• 由来する生物種: Neisseria gonorrhoeae
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 119478.78

構造登録者

Fedarovich, A.,Davies, C. (登録日: 2016-07-08, 公開日: 2017-03-01, 最終更新日: 2024-10-23)

主引用文献

Tomberg, J.,Fedarovich, A.,Vincent, L.R.,Jerse, A.E.,Unemo, M.,Davies, C.,Nicholas, R.A.
Alanine 501 Mutations in Penicillin-Binding Protein 2 from Neisseria gonorrhoeae: Structure, Mechanism, and Effects on Cephalosporin Resistance and Biological Fitness.
Biochemistry, 56:1140-1150, 2017

PubMed Abstract: Resistance of Neisseria gonorrhoeae to expanded-spectrum cephalosporins such as ceftriaxone and cefixime has increased markedly in the past decade. The primary cephalosporin resistance determinant is a mutated penA gene, which encodes the essential peptidoglycan transpeptidase, penicillin-binding protein 2 (PBP2). Decreased susceptibility and resistance can be conferred by mosaic penA alleles containing upward of 60 amino acid changes relative to wild-type PBP2, or by nonmosaic alleles with relatively few mutations, the most important of which occurs at Ala501 located near the active site of PBP2. Recently, fully cefixime- and ceftriaxone-resistant clinical isolates that harbored a mosaic penA allele with an A501P mutation were identified. To examine the potential of mutations at Ala501 to increase resistance to expanded-spectrum cephalosporins, we randomized codon 501 in a mosaic penA allele and transformed N. gonorrhoeae to increased cefixime resistance. Interestingly, only five substitutions of Ala501 (A501V, A501T, A501P, A501R, and A501S) that increased resistance and preserved essential transpeptidase function were isolated. To understand their structural implications, these mutations were introduced into the nonmosaic PBP2-6140CT, which contains four C-terminal mutations present in PBP2 from the penicillin-resistant strain FA6140. The crystal structure of PBP2-6140CT-A501T was determined and revealed ordering of a loop near the active site and a new hydrogen bond involving Thr501 that connects the loop and the SxxK conserved active site motif. The structure suggests that increased rigidity in the active site region is a mechanism for cephalosporin resistance mediated by Ala501 mutations in PBP2.

PubMed: 28145684
DOI: 10.1021/acs.biochem.6b01030

実験手法

X-RAY DIFFRACTION (2.4 Å)