Crystal structures of the class D β-lactamase OXA-13 in the native form and in complex with meropenem

Pernot, L.; Frénois, Fredéric; Rybkine, Tania; L’Hermite, Guillaume; Pétrella, S.; Delettré, J.; Jarlier, Vincent; Collatz, E.; Sougakoff, Wladimir

doi:10.1006/jmbi.2001.4805

Cited by 61 publications

(104 citation statements)

References 54 publications

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“…The structure displays an overall organization very similar to that found in other class A, C, and D ␤-lactamases (15)(16)(17)(18)(19), as well as in other DD-transpeptidases (20,21). For example, the rms difference in the atomic positions for the common C ␣ residues of OXA-24 is 1.3 Å (225 C ␣ atoms) when compared with that of the oxacillinase class D ␤-lactamase OXA-10 (17, 19) and 2.4 Å (169 C ␣ atoms) when compared with a typical class A enzyme (Escherichia coli TEM-1; ref.…”

Section: Resultssupporting

confidence: 58%

“…The structures of class A and C ␤-lactamases, as well as that of the prototypic class D oxacillinases (OXA-1, OXA-2, OXA-10, and OXA-13), have been determined (15)(16)(17)(18)(19). Comparison of these active-serine enzymes has identified similarities in their overall folding involving one helical and one mixed ␣ /␤ domain and three highly conserved active site elements in a cleft located between the two domains that are involved in hydrolysis.…”

mentioning

confidence: 99%

“…Comparison of these active-serine enzymes has identified similarities in their overall folding involving one helical and one mixed ␣ /␤ domain and three highly conserved active site elements in a cleft located between the two domains that are involved in hydrolysis. Despite these similarities, important differences both adjacent to the active site and in the omega loop region may establish their specific hydrolytic profiles (17,18). However, no structures of a class D carbapenemase are currently available.…”

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confidence: 99%

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Crystal structure of the carbapenemase OXA-24 reveals insights into the mechanism of carbapenem hydrolysis

Santillana

Beceiro

Bou

et al. 2007

Proc. Natl. Acad. Sci. U.S.A.

107

162

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Combating bacterial resistance to ␤-lactams, the most widely used antibiotics, is an emergent and clinically important challenge. OXA-24 is a class D ␤-lactamase isolated from a multiresistant epidemic clinical strain of Acinetobacter baumannii. We have investigated how OXA-24 specifically hydrolyzes the last resort carbapenem antibiotic, and we have determined the crystal structure of OXA-24 at a resolution of 2.5 Å. The structure shows that the carbapenem's substrate specificity is determined by a hydrophobic barrier that is established through the specific arrangement of the Tyr-112 and Met-223 side chains, which define a tunnel-like entrance to the active site. The importance of these residues was further confirmed by mutagenesis studies. Biochemical and microbiological analyses of specific point mutants selected on the basis of structural criteria significantly reduced the catalytic efficiency (kcat/Km) against carbapenems, whereas the specificity for oxacillin was noticeably increased. This is the previously unrecognized crystal structure that has been obtained for a class D carbapenemase enzyme. Accordingly, this information may help to improve the development of effective new drugs to combat ␤-lactam resistance. More specifically, it may help to overcome carbapenem resistance in A. baumannii, probably one of the most worrying infectious threats in hospitals worldwide.␤-lactamases ͉ carbapenem resistance ͉ enzyme mechanism ͉ protein crystallography ͉ Acinetobacter baumannii

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Section: Resultssupporting

confidence: 58%

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confidence: 99%

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confidence: 99%

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Crystal structure of the carbapenemase OXA-24 reveals insights into the mechanism of carbapenem hydrolysis

Santillana

Beceiro

Bou

et al. 2007

Proc. Natl. Acad. Sci. U.S.A.

107

162

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“…In either case, carboxylation of Lys 392 was not observed at any contour level in our electron density maps. In the class D ␤-lactamase structures, the non-carboxylated Lys 70 (corresponding to Lys 392 in BlaR1) seems to encourage an "inactive" conformation of Ser 115 (Ser 437 in BlaR1), in which the serine hydroxyl (presumed to shuttle a proton from the carboxylated lysine to the leaving group nitrogen of the ␤-lactam substrate) is somewhat displaced from its typical position in the active site (49,52). In contrast with many of the non-carboxylated structures of the class D ␤-lactamases, Kerff et al (19) noted that the active site of the apo form of the B. licheniformis BlaR1 sensor domain (which also lacks a carboxylated active-site lysine despite the fact that the crystals were grown at pH 7.0) closely resembles the "active" conformation.…”

Section: Role Of Lys 392 As the General Base In Acylation-mentioning

confidence: 99%

“…Mutation of either Pro 49 or Pro 52 to alanine in the L2 loop was observed to negate inducibility of ␤-lactamase expression (Table II). In fact, ␤-lactamase activity was not even detectable in the proline mutants because the cephaloridine concentrations were virtually identical to those of the ␤-lactamase-negative RN4220 control.…”

Section: Figmentioning

confidence: 99%

Crystal Structures of the Apo and Penicillin-acylated Forms of the BlaR1 β-Lactam Sensor of Staphylococcus aureus

Wilke

Hills

Zhang

et al. 2004

Journal of Biological Chemistry

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Staphylococcus aureus is among the most prevalent and antibiotic-resistant of pathogenic bacteria. The resistance of S. aureus to prototypal ␤-lactam antibiotics is conferred by two mechanisms: (i) secretion of hydrolytic ␤-lactamase enzymes and (ii) production of ␤-lactam-insensitive penicillin-binding proteins (PBP2a). Despite their distinct modes of resistance, expression of these proteins is controlled by similar regulation systems, including a repressor (BlaI/MecI) and a multidomain transmembrane receptor (BlaR1/MecR1). Resistance is triggered in response to a covalent binding event between a ␤-lactam antibiotic and the extracellular sensor domain of BlaR1/MecR1 by transduction of the binding signal to an intracellular protease domain capable of repressor inactivation. This study describes the first crystal structures of the sensor domain of BlaR1 (BlaR S ) from S. aureus in both the apo and penicillin-acylated forms. The structures show that the sensor domain resembles the ␤-lactam-hydrolyzing class D ␤-lactamases, but is rendered a penicillin-binding protein due to the formation of a very stable acyl-enzyme. Surprisingly, conformational changes upon penicillin binding were not observed in our structures, supporting the hypothesis that transduction of the antibiotic-binding signal into the cytosol is mediated by additional intramolecular interactions of the sensor domain with an adjacent extracellular loop in BlaR1.The evolution and dissemination of antibiotic resistance in pathogenic bacteria are a growing concern. Many of the antimicrobial "wonder drugs" society has come to rely on for the treatment of bacterial infections have been neutralized by new strains equipped with a variety of molecular countermeasures. Methicillin-resistant Staphylococcus aureus is a prominent example of this (1-3). Although ␤-lactam antibiotics such as penicillin have long been the drugs of choice for infections of S. aureus, current therapies for methicillin-resistant S. aureus infections now depend on distinct antibiotic classes such as glycopeptides to counter the increased resistance to penicillin and its derivatives. The recent emergence of glycopeptide-resistant strains of S. aureus (4, 5) underscores the need not only for the development of novel therapeutics, but for better understanding of the molecular mechanisms involved in antibiotic resistance.␤-Lactam antibiotics kill bacteria by inhibiting the cell wall transpeptidases (also known as penicillin-binding proteins (PBPs) 1 ) that are responsible for the essential cross-linking of peptidoglycan during synthesis of the rigid bacterial cell wall. Resistance to ␤-lactam antibiotics in S. aureus can be conferred by two mechanisms. In one case, ␤-lactamase enzymes are secreted from the bacterium to hydrolytically inactivate ␤-lactam antibiotics. Resistance of this form is encoded by the blaZ gene and is carried by Ͼ95% of S. aureus isolates (3). The second resistance mechanism is expression of PBP2a, a cell wall transpeptidase with broad-spectrum insensitivity to ␤-lacta...

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β‐Lactam Antibiotics

Testero

Fisher

Mobashery

2010

Burger's Medicinal Chemistry and Drug Discovery

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The β‐lactam classes of antibacterials are preeminent in the treatment of bacterial infection due to their unparalleled clinical efficacy and clinical safety. Following the discovery of the penicillins, successive β‐lactam drug discovery has added the cephalosporin, penem cephamycin, clavulanate, monobactam, nocardicin, and carbapenem subclasses. The driving force behind much of this era of discovery is the staggering ability of pathogenic bacteria to adapt previous generations of the β‐lactam by the acquisition and expression of resistance mechanisms. Although many factors contribute to β‐lactam resistance, alterations to the molecular targets of the β‐lactams (the penicillin binding proteins) and the use of enzymes (the β‐lactamases) capable of the hydrolytic deactivation of the β‐lactams are paramount. This review traces the historical development of β‐lactam drug discovery, with emphasis on the most recent progress in the medicinal chemistry, biochemistry, and microbiology of the β‐lactams leading to the discovery of new generation β‐lactam antibacterials effective against the Gram‐negative and ‐positive bacterial pathogens of current medical concern.

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Crystal structures of the class D β-lactamase OXA-13 in the native form and in complex with meropenem

Cited by 61 publications

References 54 publications

Crystal structure of the carbapenemase OXA-24 reveals insights into the mechanism of carbapenem hydrolysis

Crystal structure of the carbapenemase OXA-24 reveals insights into the mechanism of carbapenem hydrolysis

Crystal Structures of the Apo and Penicillin-acylated Forms of the BlaR1 β-Lactam Sensor of Staphylococcus aureus

β‐Lactam Antibiotics

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