Catalytic properties of class A β-lactamases: efficiency and diversity

Matagne, André; Lamotte‐Brasseur, Josette; Frère, Jean‐Marie

doi:10.1042/bj3300581

Cited by 338 publications

(321 citation statements)

References 200 publications

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“…Molecular comparisons of the various class A enzymes were facilitated by the use of a standard numbering scheme, as indicated by the label "ABL" (for class A ␤-lactamase) (96). Several years later, Matagne et al confirmed these findings and updated the list of residues that are involved in the catalytic mechanism and/or in substrate binding (12). Such characteristic residues have also recently been identified in 67 putative ␤-lactamases (97).…”

Section: Primary Structure/sequence Analysissupporting

confidence: 50%

“…This second motif was also observed in TEM-1 produced by Gramnegative bacteria (96). Given the narrow spectrum of activity of such enzymes, no amino acid substitution for expanded-spectrum resistance or resistance to inhibitors could be identified (12,114,115). Finally, the deletion of two residues between Ser218 and Ile221, in enzymes produced by several mycobacteria such as M. fortuitum, was found to be unique to the class A ␤-lactamases of Gram-positive bacteria (Fig.…”

Section: Enzymes Produced By Gram-positive Bacteriamentioning

confidence: 89%

“…As early as 1980, Ambler proposed that the following motifs are characteristic for class A ␤-lactamases: 70SerxxLys (where x's represent variable amino acids), 130SerAspAsn (the "SDN" motif), and 234LysThr/SerGly (the "KTG" motif) (6). The Glu166 residue in the ⍀-loop was found to be critical for the fast hydrolysis of penicillins and for distinguishing between class A ␤-lactamases and other serine proteins such as DD-peptidases or PBPs (11,12). Moreover, it was recently confirmed that the 166AspxxLysAsn motif (the "ExxLN" motif) is crucial for the definition of this molecular class (13).…”

Section: Phylogeny Analysismentioning

confidence: 99%

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A Structure-Based Classification of Class A β-Lactamases, a Broadly Diverse Family of Enzymes

Slama²,

et al. 2016

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SUMMARYFor medical biologists, sequencing has become a commonplace technique to support diagnosis. Rapid changes in this field have led to the generation of large amounts of data, which are not always correctly listed in databases. This is particularly true for data concerning class A β-lactamases, a group of key antibiotic resistance enzymes produced by bacteria. Many genomes have been reported to contain putative β-lactamase genes, which can be compared with representative types. We analyzed several hundred amino acid sequences of class A β-lactamase enzymes for phylogenic relationships, the presence of specific residues, and cluster patterns. A clear distinction was first made between dd-peptidases and class A enzymes based on a small number of residues (S70, K73, P107, 130SDN132, G144, E166, 234K/R, 235T/S, and 236G [Ambler numbering]). Other residues clearly separated two main branches, which we named subclasses A1 and A2. Various clusters were identified on the major branch (subclass A1) on the basis of signature residues associated with catalytic properties (e.g., limited-spectrum β-lactamases, extended-spectrum β-lactamases, and carbapenemases). For subclass A2 enzymes (e.g., CfxA, CIA-1, CME-1, PER-1, and VEB-1), 43 conserved residues were characterized, and several significant insertions were detected. This diversity in the amino acid sequences of β-lactamases must be taken into account to ensure that new enzymes are accurately identified. However, with the exception of PER types, this diversity is poorly represented in existing X-ray crystallographic data.

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Section: Primary Structure/sequence Analysissupporting

confidence: 50%

Section: Enzymes Produced By Gram-positive Bacteriamentioning

confidence: 89%

Section: Phylogeny Analysismentioning

confidence: 99%

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A Structure-Based Classification of Class A β-Lactamases, a Broadly Diverse Family of Enzymes

Slama²,

et al. 2016

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“…The class A ␤-lactamase TEM-1 presents a significant clinical antibiotic resistance problem because of mutations that alter the substrate specificity to allow hydrolysis of extended spectrum cephalosporins or provide for inhibitor resistance (3)(4)(5)(6)(7)(8). The serine ␤-lactamase mechanism is similar to that of serine proteases (44), in that a proton is abstracted from the side chain oxygen of serine that acts a nucleophile and attacks the amide bond of the ␤-lactam ring (14,15,(17)(18)(19)(20)(21)24).…”

Section: Discussionmentioning

confidence: 99%

Structural and Biochemical Evidence That a TEM-1 β-Lactamase N170G Active Site Mutant Acts via Substrate-assisted Catalysis

Brown

Shanker

Prasad

et al. 2009

Journal of Biological Chemistry

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TEM-1 ␤-lactamase is the most common plasmid-encoded166 and the hydrolytic water. The goal of this study was to examine the importance of Asn 170 for catalysis and substrate specificity of ␤-lactam antibiotic hydrolysis. The codon for position 170 was randomized to create a library containing all 20 possible amino acids. The random library was introduced into Escherichia coli, and functional clones were selected on agar plates containing ampicillin. DNA sequencing of the functional clones revealed that only asparagine (wild type) and glycine at this position are consistent with wild-type function. The determination of kinetic parameters for several substrates revealed that the N170G mutant is very efficient at hydrolyzing substrates that contain a primary amine in the antibiotic R-group that would be close to the Asn 170 side chain in the acyl-intermediate. In addition, the x-ray structure of the N170G enzyme indicated that the position of an active site water important for deacylation is altered compared with the wildtype enzyme. Taken together, the results suggest the N170G TEM-1 enzyme hydrolyzes ampicillin efficiently because of substrate-assisted catalysis where the primary amine of the ampicillin R-group positions the hydrolytic water and allows for efficient deacylation.

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“…cleaved, inactive ␤-lactam) (38). The formation of the enzymatically inactive (covalent) acyl-enzyme complex (E-I) is known as the acylation step.…”

Section: Comparison Of Tp47 Withmentioning

confidence: 99%

Crystal Structure of the 47-kDa Lipoprotein of Treponema pallidum Reveals a Novel Penicillin-binding Protein

Deka¹,

Machius²,

Norgard³

et al. 2002

Journal of Biological Chemistry

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Syphilis is a complex sexually transmitted disease caused by the spirochetal bacterium Treponema pallidum. T. pallidum has remained exquisitely sensitive to penicillin, but the mode of action and lethal targets for ␤-lactams are still unknown. We previously identified the T. pallidum 47-kDa lipoprotein (Tp47) as a penicillin-binding protein (PBP). Tp47 contains three hypothetical consensus motifs (SVTK, TEN, and KTG) that typically form the active center of other PBPs. Yet, in this study, mutations of key amino acids within these motifs failed to abolish the penicillin binding activity of Tp47. The crystal structure of Tp47 at a resolution of 1.95 Å revealed a fold different from any other known PBP; Tp47 is predominantly ␤-sheet, in contrast to the ␣/␤-fold common to other PBPs. It comprises four distinct domains: two complex ␤-sheet-containing N-terminal domains and two C-terminal domains that adopt immunoglobulin-like folds. The three hypothetical PBP signature motifs do not come together to form a typical PBP active site. Furthermore, Tp47 is unusual in that it displays ␤-lactamase activity (k cat for penicillin ‫؍‬ 271 ؎ 6 s ؊1 ), a feature that hindered attempts to identify the active site in Tp47 by co-crystallization and mass spectrometric techniques. Taken together, Tp47 does not fit the classical structural and mechanistic paradigms for PBPs, and thus Tp47 appears to represent a new class of PBP.

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Catalytic properties of class A β-lactamases: efficiency and diversity

Cited by 338 publications

References 200 publications

A Structure-Based Classification of Class A β-Lactamases, a Broadly Diverse Family of Enzymes

A Structure-Based Classification of Class A β-Lactamases, a Broadly Diverse Family of Enzymes

Structural and Biochemical Evidence That a TEM-1 β-Lactamase N170G Active Site Mutant Acts via Substrate-assisted Catalysis

Crystal Structure of the 47-kDa Lipoprotein of Treponema pallidum Reveals a Novel Penicillin-binding Protein

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