Atomic‐Resolution Structure of a Class C β‐Lactamase and Its Complex with Avibactam

Pozzi, Cecilia; Pisa, F. Di; Luca, Filomena De; Benvenuti, Manuela; Docquier, Jean Denis; Mangani, Stefano

doi:10.1002/cmdc.201800213

Cited by 15 publications

(17 citation statements)

References 46 publications

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“…[5,6]. Class C β-lactamases play a role in the resistance of these pathogens to cephalosporins, cephamycins, and carbapenems, and are not inhibited by clavulanic acid [7,8]. The molecular structure of class C β-lactamases consists of two domains of different sizes.…”

Section: Introductionmentioning

confidence: 99%

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Molecular Characterization of a Novel Family VIII Esterase with β-Lactamase Activity (PsEstA) from Paenibacillus sp.

et al. 2019

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Molecular information about family VIII esterases, which have similarities with class C β-lactamases and penicillin-binding proteins, remains largely unknown. In this study, a novel family VIII esterase with β-lactamase activity (PsEstA) from Paenibacillus sp. was characterized using several biochemical and biophysical methods. PsEstA was effective on a broad range of substrates including tertiary butyl acetate, glyceryl tributyrate, glucose pentaacetate, olive oil, and p-nitrophenyl esters. Additionally, PsEstA hydrolyzed nitrocefin, cefotaxime, and 7-aminocephalosporanic acid. Interestingly, two forms of immobilized PsEstA (CLEAs-PsEstA and mCLEAs-PsEstA) showed high recycling property and enhanced stability, but hybrid nanoflowers (hNFs) of PsEstA require improvement. This study provides a molecular understanding of substrate specificities, catalytic regulation, and immobilization of PsEstA, which can be efficiently used in biotechnological applications.

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Section: Introductionmentioning

confidence: 99%

“…The molecular structure of class C β-lactamases consists of two domains of different sizes. The bigger domain contains a central antiparallel beta sheet flanked by alpha helices at each side, while the smaller domain is entirely composed of alpha helices and contains the catalytic serine residue [7,[9][10][11].…”

Section: Introductionmentioning

confidence: 99%

Molecular Characterization of a Novel Family VIII Esterase with β-Lactamase Activity (PsEstA) from Paenibacillus sp.

et al. 2019

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“…While both IDCs display a resistance pattern consistent with that of an AmpC, IDC-1 causes much higher resistance levels than IDC-2 in an isogenic background. All three conserved sequence motifs necessary for class C β-lactamase activity are identical in IDC-1 and IDC-2 [76,77]. However, 18 amino acid changes were detected outside of the conserved motifs, of which one or several might be responsible for the observed differences in activity.…”

Section: Discussionmentioning

confidence: 94%

A Novel, Integron-Regulated, Class C β-Lactamase

et al. 2020

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AmpC-type β-lactamases severely impair treatment of many bacterial infections, due to their broad spectrum (they hydrolyze virtually all β-lactams, except fourth-generation cephalosporins and carbapenems) and the increasing incidence of plasmid-mediated versions. The original chromosomal AmpCs are often tightly regulated, and their expression is induced in response to exposure to β-lactams. Regulation of mobile ampC expression is in many cases less controlled, giving rise to constitutively resistant strains with increased potential for development or acquisition of additional resistances. We present here the identification of two integron-encoded ampC genes, blaIDC-1 and blaIDC-2 (integron-derived cephalosporinase), with less than 85% amino acid sequence identity to any previously annotated AmpC. While their resistance pattern identifies them as class C β-lactamases, their low isoelectric point (pI) values make differentiation from other β-lactamases by isoelectric focusing impossible. To the best of our knowledge, this is the first evidence of an ampC gene cassette within a class 1 integron, providing a mobile context with profound potential for transfer and spread into clinics. It also allows bacteria to adapt expression levels, and thus reduce fitness costs, e.g., by cassette-reshuffling. Analyses of public metagenomes, including sewage metagenomes, show that the discovered ampCs are primarily found in Asian countries.

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“…While the interactions of avibactam with a wide variety of SBLs have been studied (e.g. (24)(25)(26)(27)(28)(29)(30)), structural information on the binding of other DBOs is more limited (16,19,20,(31)(32)(33).…”

Section: Downloaded Frommentioning

confidence: 99%

Structural Investigations of the Inhibition of Escherichia coli AmpC β-Lactamase by Diazabicyclooctanes

Lang

Leissing

Page

et al. 2021

Antimicrob Agents Chemother

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β-Lactam antibiotics are presently the most important treatments for infections by pathogenic Escherichia coli, but their use is increasingly compromised by β-lactamases, including the chromosomally encoded class C AmpC serine-β-lactamases (SBL). The diazabicyclooctane (DBO) avibactam is a potent AmpC inhibitor; the clinical success of avibactam combined with ceftazidime has stimulated efforts to optimise the DBO core. We report kinetic and structural studies, including four high resolution crystal structures, concerning inhibition of the AmpC serine-β-lactamase from E. coli (AmpCEC) by clinically relevant DBO-based inhibitors: avibactam, relebactam, nacubactam, and zidebactam. Kinetic analyses and mass spectrometry-based assays were used to study their mechanisms of AmpCEC inhibition. The results reveal that, under our assay conditions, zidebactam manifests increased potency (Kiapp 0.69 μM) against AmpCEC compared to the other DBOs (Kiapp 5.0-7.4 μM) due to an ∼ 10 fold accelerated carbamoylation-rate. However, zidebactam also has an accelerated off-rate and with sufficient preincubation time all the DBOs manifest similar potencies. Crystallographic analyses indicate a greater conformational freedom of the AmpCEC-zidebactam carbamoyl-complex compared to those for the other DBOs. The results suggest carbamoyl-complex lifetime should be a consideration in development of DBO-based SBL inhibitors for the clinically important class C SBLs.

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Atomic‐Resolution Structure of a Class C β‐Lactamase and Its Complex with Avibactam

Cited by 15 publications

References 46 publications

Molecular Characterization of a Novel Family VIII Esterase with β-Lactamase Activity (PsEstA) from Paenibacillus sp.

Molecular Characterization of a Novel Family VIII Esterase with β-Lactamase Activity (PsEstA) from Paenibacillus sp.

A Novel, Integron-Regulated, Class C β-Lactamase

Structural Investigations of the Inhibition of Escherichia coli AmpC β-Lactamase by Diazabicyclooctanes

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