Biochemical characterization of IMP-30, a metallo-β-lactamase with enhanced activity toward ceftazidime.
Abstract
IMP-type enzymes constitute a clinically important family of metallo-β-lactamases that has grown dramatically in the past decade to its current 45 known members. Here, we report the biochemical characterization of IMP-30 in comparison to IMP-1, from which it deviates by a single E59K mutation. Kinetics, MIC assays, docking, and molecular dynamics simulations support a scenario in which Lys59 interacts with the ceftazidime R1 group, resulting in increased water access and enhanced turnover and MIC of ceftazidime.
Submission Details
ID: Z5J9b5KA4
Submitter: Peter Oelschlaeger
Submission Date: Aug. 20, 2019, 11:12 a.m.
Version: 1
Publication Details
Pegg KM;Liu EM;Lacuran AE;Oelschlaeger P,Antimicrob Agents Chemother (2013) Biochemical characterization of IMP-30, a metallo-β-lactamase with enhanced activity toward ceftazidime. PMID:23836186Additional Information
Explanation of amino acid numbering: Mutant E23K of IMP-1 is a wild-type enzyme called IMP-30. According to the standard numbering scheme (PMID 15215079) its mutation is E59K. ND indicates not detectable (used for kinetic constants with aztreonam).
Study Summary
| Number of data points | 180 |
| Proteins | Metallo-beta-lactamase type 2 |
| Unique complexes | 20 |
| Assays/Quantities/Protocols | Experimental Assay: kcat ; Experimental Assay: Resistance ; Experimental Assay: MIC relative ; Experimental Assay: MIC absolute ; Experimental Assay: kcat/Km ; Experimental Assay: Km ; Derived Quantity: SD of kcat/Km ; Derived Quantity: SD of Km ; Derived Quantity: SD of kcat |
| Libraries | Activity data for IMP-1 and IMP-30 (IMP-1-E59K) |
Structure view and single mutant data analysis
Study data
| 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 |
|---|---|---|---|
| 1DD6 | 1999-11-08T00:00:00+0000 | 2.0 | IMP-1 METALLO BETA-LACTAMASE FROM PSEUDOMONAS AERUGINOSA IN COMPLEX WITH A MERCAPTOCARBOXYLATE INHIBITOR |
| 1VGN | 2004-04-27T00:00:00+0000 | 2.63 | Structure-based design of the irreversible inhibitors to metallo--lactamase (IMP-1) |
| 1WUO | 2004-12-08T00:00:00+0000 | 2.01 | Crystal structure of metallo-beta-lactamase IMP-1 mutant (D81A) |
| 1WUP | 2004-12-08T00:00:00+0000 | 3.0 | Crystal structure of metallo-beta-lactamase IMP-1 mutant (D81E) |
| 2DOO | 2006-05-01T00:00:00+0000 | 2.43 | The structure of IMP-1 complexed with the detecting reagent (DansylC4SH) by a fluorescent probe |
| 4C1F | 2013-08-12T00:00:00+0000 | 2.01 | Crystal structure of the metallo-beta-lactamase IMP-1 with L-captopril |
| 4C1G | 2013-08-12T00:00:00+0000 | 1.71 | Crystal structure of the metallo-beta-lactamase IMP-1 with D-captopril |
| 5EV6 | 2015-11-19T00:00:00+0000 | 1.98 | Crystal structure of the native, di-zinc metallo-beta-lactamase IMP-1 |
| 5EV8 | 2015-11-19T00:00:00+0000 | 2.3 | Crystal structure of the metallo-beta-lactamase IMP-1 in complex with the bisthiazolidine inhibitor D-CS319 |
| 5EWA | 2015-11-20T00:00:00+0000 | 2.3 | Crystal structure of the metallo-beta-lactamase IMP-1 in complex with the bisthiazolidine inhibitor L-VC26 |
Relevant UniProtKB Entries
| Percent Identity | Matching Chains | Protein | Accession | Entry Name |
|---|---|---|---|---|
| 100.0 | A | Metallo-beta-lactamase type 2 | P52699 | BLAB_SERMA |