Mechanistic study of the hydrolysis of nitrocefin mediated by B.cereus metallo-β-lactamase

Rasia, Rodolfo M.; Vila, A.J.

doi:10.3998/ark.5550190.0004.a47

Cited by 18 publications

(26 citation statements)

References 17 publications

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“…Behavior varies widely across the B1 MBL subclass, as summarized in Table 5 . 24 − 27 , 48 , 49 , 55 , 57 − 60 To support this, we include two reports on CcrA here. One is based on an early study by Benkovic and co-workers that reported an optical titration consistent with distributed Co(II) binding.…”

Section: Discussionmentioning

confidence: 99%

Biochemical, Mechanistic, and Spectroscopic Characterization of Metallo-β-lactamase VIM-2

et al. 2014

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This study examines metal binding to metallo-β-lactamase VIM-2, demonstrating the first successful preparation of a Co(II)-substituted VIM-2 analogue. Spectroscopic studies of the half- and fully metal loaded enzymes show that both Zn(II) and Co(II) bind cooperatively, where the major species present, regardless of stoichiometry, are apo- and di-Zn (or di-Co) enzymes. We determined the di-Zn VIM-2 structure to a resolution of 1.55 Å, and this structure supports results from spectroscopic studies. Kinetics, both steady-state and pre-steady-state, show that VIM-2 utilizes a mechanism that proceeds through a very short-lived anionic intermediate when chromacef is used as the substrate. Comparison with other B1 enzymes shows that those that bind Zn(II) cooperatively are better poised to protonate the intermediate on its formation, compared to those that bind Zn(II) non-cooperatively, which uniformly build up substantial amounts of the intermediate.

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Section: Discussionmentioning

confidence: 99%

Biochemical, Mechanistic, and Spectroscopic Characterization of Metallo-β-lactamase VIM-2

et al. 2014

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“…In contrast to these results, no accumulation of an anionic intermediate could be detected in reactions catalysed by BCII and the rate of substrate depletion and product formation are similar [ 55 ]. However, for the BCII-catalysed hydrolysis under cryoenzymological conditions, a slightly red-shifted intermediate (440 nm) was observed, which was suggested to be a nitrocefin-like tetrahedral intermediate [ 56 ].…”

Section: Metallo β -Lactamasesmentioning

confidence: 91%

“…Based on these observations it was concluded that both the replacement of the arginine residue and the introduction of the second metal-ion are evolutionary tools for accelerating C–N bond fission in MBL catalysed hydrolysis of β -lactams. However, an important role in lowering the energy barrier for breaking the C–N bond must come from other enzyme structural rearrangements, since the reciprocal, that is, the sole replacement of the arginine by a cysteine residue and the insertion of a second zinc-ion in BCII, only marginally (two fold) improves the catalytic activity [ 55 ].…”

Section: Metallo β -Lactamasesmentioning

confidence: 99%

The Mechanisms of Catalysis by Metallo β‐Lactamases

Page

Badarau

2008

Bioinorganic Chemistry and Applications

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Class B β-lactamases or metallo-β-lactamases (MBLs) require zinc ions to catalyse the hydrolysis of β-lactam antibiotics such as penicillins, cephalosporins, carbapenems, and cephamycins. There are no clinically useful inhibitors against MBLs which are responsible for the resistance of some bacteria to antibiotics. There are two metal-ion binding sites that have different zinc ligands but the exact roles of the metal-ion remain controversial, and distinguishing between their relative importance is complex. The metal-ion can act as a Lewis acid by co-ordination to the β-lactam carbonyl oxygen to facilitate nucleophilic attack and stabilise the negative charge developed on this oxygen in the tetrahedral intermediate anion. The metal-ion also lowers the pKa of the directly co-ordinated water molecule so that the metal-bound hydroxide ion is a better nucleophile than water and is used to attack the β-lactam carbonyl carbon. An intrinsic property of binuclear metallo hydrolytic enzymes that depend on a metal-bound water both as the attacking nucleophile and as a ligand for the second metal-ion is that this water molecule, which is consumed during hydrolysis of the substrate, has to be replaced to maintain the catalytic cycle. With MBL this is reflected in some unusual kinetic profiles.

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“…The accumulation of the nitrocefin intermediate is different among MBLs: there is barely any intermediate formed with BcII or IMP-1 . Minor perturbations, such as as second sphere mutations that change the hydrogen bond network or loop changes, can lead to large changes in the accumulation of this and other intermediates .…”

Section: Catalytic Mechanism Of Mblsmentioning

confidence: 99%

Metallo-β-lactamases in the Age of Multidrug Resistance: From Structure and Mechanism to Evolution, Dissemination, and Inhibitor Design

2021

Self Cite

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Antimicrobial resistance is one of the major problems in current practical medicine. The spread of genes coding for resistance determinants among bacteria challenges the use of approved antibiotics, narrowing the options for treatment. Resistance to carbapenems, last resort antibiotics, is a major concern. Metallo-β-lactamases (MBLs) hydrolyze carbapenems, penicillins, and cephalosporins, becoming central to this problem. These enzymes diverge with respect to serine-β-lactamases by exhibiting a different fold, active site, and catalytic features. Elucidating their catalytic mechanism has been a big challenge in the field that has limited the development of useful inhibitors. This review covers exhaustively the details of the active-site chemistries, the diversity of MBL alleles, the catalytic mechanism against different substrates, and how this information has helped developing inhibitors. We also discuss here different aspects critical to understand the success of MBLs in conferring resistance: the molecular determinants of their dissemination, their cell physiology, from the biogenesis to the processing involved in the transit to the periplasm, and the uptake of the Zn(II) ions upon metal starvation conditions, such as those encountered during an infection. In this regard, the chemical, biochemical and microbiological aspects provide an integrative view of the current knowledge of MBLs.

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Mechanistic study of the hydrolysis of nitrocefin mediated by B.cereus metallo-β-lactamase

Cited by 18 publications

References 17 publications

Biochemical, Mechanistic, and Spectroscopic Characterization of Metallo-β-lactamase VIM-2

Biochemical, Mechanistic, and Spectroscopic Characterization of Metallo-β-lactamase VIM-2

The Mechanisms of Catalysis by Metallo β‐Lactamases

Metallo-β-lactamases in the Age of Multidrug Resistance: From Structure and Mechanism to Evolution, Dissemination, and Inhibitor Design

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