A Kinetic Study of the Replacement by Site Saturation Mutagenesis of Residue 119 in NDM-1 Metallo-β-Lactamase.

Abstract

New Delhi metallo-β-lactamase 1 (NDM-1) is a subclass B1 metallo-β-lactamase that exhibits a broad spectrum of activity against β-lactam antibiotics. Here we report the kinetic study of 6 Q119X variants obtained by site-directed mutagenesis of NDM-1. All Q119X variants were able to hydrolyze carbapenems, penicillins and first-, second-, third-, and fourth-generation cephalosporins very efficiently. In particular, Q119E, Q119Y, Q119V, and Q119K mutants showed improvements in kcat/Km values for penicillins, compared with NDM-1. The catalytic efficiencies of the Q119K variant for benzylpenicillin and carbenicillin were about 65- and 70-fold higher, respectively, than those of NDM-1. The Q119K and Q119Y enzymes had kcat/Km values for ceftazidime about 25- and 89-fold higher, respectively, than that of NDM-1.

Submission Details

ID: CWvASvaZ3

Submitter: Shu-Ching Ou

Submission Date: March 20, 2019, 3:27 p.m.

Version: 1

Publication Details
Marcoccia F;Mercuri PS;Galleni M;Celenza G;Amicosante G;Perilli M,Antimicrob Agents Chemother (2018) A Kinetic Study of the Replacement by Site Saturation Mutagenesis of Residue 119 in NDM-1 Metallo-β-Lactamase. PMID:29784851
Additional Information

Study Summary

Number of data points 299
Proteins Metallo-beta-lactamase type 2
Unique complexes 117
Assays/Quantities/Protocols Experimental Assay: kcat/Km ; Experimental Assay: Km ; Experimental Assay: kcat ; Experimental Assay: MIC
Libraries Variants for NDM-1_Substrate

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
3PG4 2010-10-29T00:00:00+0000 2.0 The crystal structure of New Delhi Metallo-beta lactamase (NDM-1)
3RKJ 2011-04-18T00:00:00+0000 2.0 Crystal Structure of New Delhi Metallo-Beta-Lactamase-1 from Klebsiella pnueumoniae
3RKK 2011-04-18T00:00:00+0000 2.35 Crystal Structure of New Delhi Metallo-Beta-Lactamase-1 from Klebsiella pneumoniae
3SBL 2011-06-05T00:00:00+0000 2.31 Crystal Structure of New Delhi Metal-beta-lactamase-1 from Klebsiella pneumoniae
3SFP 2011-06-13T00:00:00+0000 2.27 Crystal Structure of the Mono-Zinc-boundform of New Delhi Metallo-beta-Lactamase-1 from Klebsiella pneumoniae
3SPU 2011-07-03T00:00:00+0000 2.1 apo NDM-1 Crystal Structure
3SRX 2011-07-07T00:00:00+0000 2.5 New Delhi Metallo-beta-Lactamase-1 Complexed with Cd
3ZR9 2011-06-15T00:00:00+0000 1.91 Structure of New Delhi Metallo-Beta-lactamase 1 (NDM-1)
4EXS 2012-04-30T00:00:00+0000 2.4 Crystal structure of NDM-1 bound to L-captopril
4EXY 2012-05-01T00:00:00+0000 1.47 Crystal structure of NDM-1 bound to ethylene glycol

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
100.0 Metallo-beta-lactamase type 2 C7C422 BLAN1_KLEPN