Irreversible inhibition of the bacterial cysteine protease-transpeptidase sortase (SrtA) by substrate-derived affinity labels

Scott, Christopher J.; McDowell, Andrew; Martin, S. Lorraine; Lynas, John F.; Vandenbroeck, Koen; Walker, Brian

doi:10.1042/bj20020602

Cited by 78 publications

(71 citation statements)

References 27 publications

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“…Isosteres of the scissile bond, i.e. threonine-glycine for sortase A, achieved inhibition when the LPXT peptides were decorated with either diazoketone or chloromethylketone (13). These peptide-derived inhibitors display favorable K i values, however their rates of inactivation are slow.…”

Section: Discussionmentioning

confidence: 99%

“…Inhibition of sortases by small molecules may therefore function as a therapeutic strategy for bacterial infections. Previous work described several sortase inhibitors, including methane-thiosulfonates (12), peptide substrate-derived affinity labels (13), natural compounds (14 -16), vinyl sulfones (17), diarylacrylonitriles (18), bis(indole) alkaloids (19), peptidomimetics (20), isoquinoline alkaloids (16), and threonine analogues (21). However, most of these compounds are either of low activity, lack specificity, or display undesirable structural features that confound therapeutic use.…”

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confidence: 99%

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Activation of Inhibitors by Sortase Triggers Irreversible Modification of the Active Site

Maresso

Kern

et al. 2007

Journal of Biological Chemistry

101

106

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Sortases anchor surface proteins to the cell wall of Gram-positive pathogens through recognition of specific motif sequences. Loss of sortase leads to large reductions in virulence, which identifies sortase as a target for the development of antibacterials. By screening 135,625 small molecules for inhibition, we report here that aryl (␤-amino)ethyl ketones inhibit sortase enzymes from staphylococci and bacilli. Inhibition of sortases occurs through an irreversible, covalent modification of their active site cysteine. Sortases specifically activate this class of molecules via ␤-elimination, generating a reactive olefin intermediate that covalently modifies the cysteine thiol. Analysis of the three-dimensional structure of Bacillus anthracis sortase B with and without inhibitor provides insights into the mechanism of inhibition and reveals binding pockets that can be exploited for drug discovery.The emergence of bacterial strains resistant to antibiotic therapy is a major public health threat (1). Of particular concern is Staphylococcus aureus, because this Gram-positive pathogen is the leading cause of infections in the bloodstream, lower respiratory tract, skin, and soft tissue in the United States (2). S. aureus strains exhibiting resistance against multiple antibiotics, such as methicillin-resistant S. aureus, are isolated in 30 -60% of community and Ͼ80% of hospital infections with this pathogen (3). Vancomycin or other glycopeptides are considered last-resort therapies for methicillin-resistant S. aureus; however, S. aureus strains with intermediate or full resistance to vancomycin can cause infections for which antimicrobial treatment may no longer be effective (4).Surface proteins of Gram-positive bacteria play important roles during pathogenesis (5). Sortases anchor these polypeptides to the bacterial cell wall envelope (6). For example, S. aureus sortase A recognizes proteins destined for the cell surface via an LPXTG motif in their C-terminal sorting signal (7). Following cleavage between the threonine and the glycine residues, an acyl-enzyme intermediate captures cleaved substrate at the active site thiol of sortase (8). Nucleophilic attack of the amino group of the peptidoglycan precursor lipid II (at the thioester intermediate resolves the acyl enzyme and forms an amide bond between the C-terminal threonine of surface protein and pentaglycine crossbridges (9). Lipid II-linked polypeptide is subsequently incorporated into the cell wall envelope of staphylococci (10). The final product of this pathway, protein linked to cell wall pentaglycine cross-bridges, is displayed on the bacterial surface and enables interactions between the pathogen and tissues of its host.Surface protein anchoring to the cell wall envelope is thought to be an essential strategy for bacterial survival during infection, because mutants lacking genes for one or more sortase enzymes are attenuated in virulence (11). Inhibition of sortases by small molecules may therefore function as a therapeutic strategy for bacterial infections. ...

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Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

Activation of Inhibitors by Sortase Triggers Irreversible Modification of the Active Site

Maresso

Kern

et al. 2007

Journal of Biological Chemistry

101

106

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“…Earlier work used in vitro (inhibition of fluorogenic substrate cleavage) and virtual screening of compound libraries to identify sortase inhibitors (15,(52)(53)(54)(55)(56). Although these studies identified both competitive and noncompetitive inhibitors (57,58), isolated compounds have not yet been shown to inhibit in vivo sortase activity in staphylococci, i.e., the cleavage of sorting signals or the assembly of surface proteins into the bacterial cell wall (54,(59)(60)(61)(62). Many of the isolated compounds diminish or block staphylococcal growth, indicating that they cannot function as selective inhibitors of S. aureus sortase (53,61,63,64).…”

Section: Discussionmentioning

confidence: 99%

Antiinfective therapy with a small molecule inhibitor of Staphylococcus aureus sortase

Zhang

Liu

Zhu

et al. 2014

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

132

145

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Methicillin-resistant Staphylococcus aureus (MRSA) is the most frequent cause of hospital-acquired infection, which manifests as surgical site infections, bacteremia, and sepsis. Due to drug-resistance, prophylaxis of MRSA infection with antibiotics frequently fails or incites nosocomial diseases such as Clostridium difficile infection. Sortase A is a transpeptidase that anchors surface proteins in the envelope of S. aureus, and sortase mutants are unable to cause bacteremia or sepsis in mice. Here we used virtual screening and optimization of inhibitor structure to identify 3-(4-pyridinyl)-6-(2-sodiumsulfonatephenyl) [1,2,4]

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“…In the first of these studies, the threonine-glycine peptide bond was substituted by moieties known to alkylate the active-site thiol of cysteine proteases. These included peptidyl-diazomethane (LPAT-CHN 2 ) and peptidyl-chloromethane (LPAT-CH 2 Cl) (176)…”

Section: Sortase a Inhibitorsmentioning

confidence: 99%

Sortases and the Art of Anchoring Proteins to the Envelopes of Gram-Positive Bacteria

Marraffini

DeDent²,

Schneewind³

2006

Microbiol Mol Biol Rev

604

600

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SUMMARY The cell wall envelopes of gram-positive bacteria represent a surface organelle that not only functions as a cytoskeletal element but also promotes interactions between bacteria and their environment. Cell wall peptidoglycan is covalently and noncovalently decorated with teichoic acids, polysaccharides, and proteins. The sum of these molecular decorations provides bacterial envelopes with species- and strain-specific properties that are ultimately responsible for bacterial virulence, interactions with host immune systems, and the development of disease symptoms or successful outcomes of infections. Surface proteins typically carry two topogenic sequences, i.e., N-terminal signal peptides and C-terminal sorting signals. Sortases catalyze a transpeptidation reaction by first cleaving a surface protein substrate at the cell wall sorting signal. The resulting acyl enzyme intermediates between sortases and their substrates are then resolved by the nucleophilic attack of amino groups, typically provided by the cell wall cross bridges of peptidoglycan precursors. The surface protein linked to peptidoglycan is then incorporated into the envelope and displayed on the microbial surface. This review focuses on the mechanisms of surface protein anchoring to the cell wall envelope by sortases and the role that these enzymes play in bacterial physiology and pathogenesis.

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Irreversible inhibition of the bacterial cysteine protease-transpeptidase sortase (SrtA) by substrate-derived affinity labels

Cited by 78 publications

References 27 publications

Activation of Inhibitors by Sortase Triggers Irreversible Modification of the Active Site

Activation of Inhibitors by Sortase Triggers Irreversible Modification of the Active Site

Antiinfective therapy with a small molecule inhibitor of Staphylococcus aureus sortase

Sortases and the Art of Anchoring Proteins to the Envelopes of Gram-Positive Bacteria

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