Elucidating the Role of Residue 67 in IMP-Type Metallo-β-Lactamase Evolution.


Antibiotic resistance in bacteria is ever changing and adapting, as once-novel β-lactam antibiotics are losing their efficacy, primarily due to the production of β-lactamases. Metallo-β-lactamases (MBLs) efficiently inactivate a broad range of β-lactam antibiotics, including carbapenems, and are often coexpressed with other antibacterial resistance factors. The rapid dissemination of MBLs and lack of novel antibacterials pose an imminent threat to global health. In an effort to better counter these resistance-conferring β-lactamases, an investigation of their natural evolution and resulting substrate specificity was employed. In this study, we elucidated the effects of different amino acid substitutions at position 67 in IMP-type MBLs on the ability to hydrolyze and confer resistance to a range of β-lactam antibiotics. Wild-type β-lactamases IMP-1 and IMP-10 and mutants IMP-1-V67A and IMP-1-V67I were characterized biophysically and biochemically, and MICs for Escherichia coli cells expressing these enzymes were determined. We found that all variants exhibited catalytic efficiencies (kcat/Km) equal to or higher than that of IMP-1 against all tested β-lactams except penicillins, against which IMP-1 and IMP-1-V67I showed the highest kcat/Km values. The substrate-specific effects of the different amino acid substitutions at position 67 are discussed in light of their side chain structures and possible interactions with the substrates. Docking calculations were employed to investigate interactions between different side chains and an inhibitor used as a β-lactam surrogate. The differences in binding affinities determined experimentally and computationally seem to be governed by hydrophobic interactions between residue 67 and the inhibitor and, by inference, the β-lactam substrates.

Submission Details

ID: rsSEfaoo

Submitter: Paulie Dang

Submission Date: Sept. 9, 2019, 10:50 a.m.

Version: 1

Publication Details
LaCuran AE;Pegg KM;Liu EM;Bethel CR;Ai N;Welsh WJ;Bonomo RA;Oelschlaeger P,Antimicrob Agents Chemother (2015) Elucidating the Role of Residue 67 in IMP-Type Metallo-β-Lactamase Evolution. PMID:26369960
Additional Information

Explanation of amino acid numbering: Residue 31 that was mutated in this study is called residue 67 according to the standard numbering scheme (PMID 15215079) and the title of the publication. The wild-type enzyme IMP-1 has the mutation V31F (V67F according to standard numbering). ND indicates not detectable.

Structure view and single mutant data analysis

Study data

No weblogo for data of varying length.
Colors: D E R H K S T N Q A V I L M F Y W C G P

Data Distribution

Studies with similar sequences (approximate matches)

Correlation with other assays (exact sequence matches)

Relevant PDB Entries

Structure ID Release Date Resolution Structure Title
6JED 2019-02-05T00:00:00+0000 1.57 Crystal structure of IMP-1 metallo-beta-lactamase in a complex with MCR
5Y5B 2017-08-08T00:00:00+0000 1.7 Crystal Structure Of IMP-1 Metallo-beta-lactamase
5HH4 2016-01-09T00:00:00+0000 2.0 Crystal structure of metallo-beta-lactamase IMP-1 in complex with a phosphonate-based inhibitor
1VGN 2004-04-27T00:00:00+0000 2.63 Structure-based design of the irreversible inhibitors to metallo--lactamase (IMP-1)
4C1F 2013-08-12T00:00:00+0000 2.01 Crystal structure of the metallo-beta-lactamase IMP-1 with L-captopril
6JKA 2019-02-28T00:00:00+0000 2.01 Crystal structure of metallo-beta-lactamse, IMP-1, in complex with a thiazole-bearing inhibitor
4C1G 2013-08-12T00:00:00+0000 1.71 Crystal structure of the metallo-beta-lactamase IMP-1 with D-captopril
6ZYS 2020-08-02T00:00:00+0000 1.87 Structure of IMP-1 with 2-Mercaptomethyl-thiazolidine D-syn-1b
6LBL 2019-11-14T00:00:00+0000 1.68 Crystal structure of IMP-1 metallo-beta-lactamase in complex with NO9 inhibitor

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
100.0 A Metallo-beta-lactamase type 2 P52699 BLAB_SERMA