PDBsum entry 5evd

Hydrolase

PDB id: 5evd

Contents

Protein chain

267 a.a.

Ligands

VC2

SO4 ×2

Metals

_ZN ×2

Waters ×271

PDB id:

5evd

Name:

Hydrolase

Title:

Crystal structure of the metallo-beta-lactamase l1 in complex with the bisthiazolidine inhibitor d-vc26

Structure:

Metallo-beta-lactamase l1. Chain: a. Synonym: beta-lactamase type ii,penicillinase. Engineered: yes

Source:

Stenotrophomonas maltophilia. Pseudomonas maltophilia. Organism_taxid: 40324. Expressed in: escherichia coli bl21(de3). Expression_system_taxid: 469008. Expression_system_variant: solu.

Resolution:

1.80Å R-factor: 0.167 R-free: 0.196

Authors:

P.Hinchliffe,J.Spencer

Key ref:

P.Hinchliffe et al. (2016). Cross-class metallo-β-lactamase inhibition by bisthiazolidines reveals multiple binding modes. Proc Natl Acad Sci U S A, 113, E3745. PubMed id: 27303030 DOI: 10.1073/pnas.1601368113

Date:

19-Nov-15 Release date: 01-Jun-16

Protein chain

P52700  (BLA1_STEMA) -  Metallo-beta-lactamase L1 type 3 from Stenotrophomonas maltophilia

Seq: 290 a.a.

Struc: 267 a.a.

Enzyme reactions

• Enzyme class: E.C.3.5.2.6 - beta-lactamase.
• Pathway: Penicillin Biosynthesis and Metabolism
• Reaction: a beta-lactam + H2O = a substituted beta-amino acid
• Cofactor: Zn(2+)

DOI no: 10.1073/pnas.1601368113

Proc Natl Acad Sci U S A 113:E3745 (2016)

PubMed id: 27303030

Cross-class metallo-β-lactamase inhibition by bisthiazolidines reveals multiple binding modes.

P.Hinchliffe, M.M.González, M.F.Mojica, J.M.González, V.Castillo, C.Saiz, M.Kosmopoulou, C.L.Tooke, L.I.Llarrull, G.Mahler, R.A.Bonomo, A.J.Vila, J.Spencer.

ABSTRACT

Metallo-β-lactamases (MBLs) hydrolyze almost all β-lactam antibiotics and are unaffected by clinically available β-lactamase inhibitors (βLIs). Active-site architecture divides MBLs into three classes (B1, B2, and B3), complicating development of βLIs effective against all enzymes. Bisthiazolidines (BTZs) are carboxylate-containing, bicyclic compounds, considered as penicillin analogs with an additional free thiol. Here, we show both l- and d-BTZ enantiomers are micromolar competitive βLIs of all MBL classes in vitro, with Kis of 6-15 µM or 36-84 µM for subclass B1 MBLs (IMP-1 and BcII, respectively), and 10-12 µM for the B3 enzyme L1. Against the B2 MBL Sfh-I, the l-BTZ enantiomers exhibit 100-fold lower Kis (0.26-0.36 µM) than d-BTZs (26-29 µM). Importantly, cell-based time-kill assays show BTZs restore β-lactam susceptibility of Escherichia coli-producing MBLs (IMP-1, Sfh-1, BcII, and GOB-18) and, significantly, an extensively drug-resistant Stenotrophomonas maltophilia clinical isolate expressing L1. BTZs therefore inhibit the full range of MBLs and potentiate β-lactam activity against producer pathogens. X-ray crystal structures reveal insights into diverse BTZ binding modes, varying with orientation of the carboxylate and thiol moieties. BTZs bind the di-zinc centers of B1 (IMP-1; BcII) and B3 (L1) MBLs via the free thiol, but orient differently depending upon stereochemistry. In contrast, the l-BTZ carboxylate dominates interactions with the monozinc B2 MBL Sfh-I, with the thiol uninvolved. d-BTZ complexes most closely resemble β-lactam binding to B1 MBLs, but feature an unprecedented disruption of the D120-zinc interaction. Cross-class MBL inhibition therefore arises from the unexpected versatility of BTZ binding.