Corrections - Simple Alkanethiol Groups for Temporary Blocking of Sulfhydryl Groups of Enzymes

Smith, Daniel; Maggio, E; Kenyon, George L.

doi:10.1021/bi00685a601

Cited by 30 publications

(42 citation statements)

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“…4). This activity appears to be specific for sortase, as it can, at least in part, be inhibited by preincubation of staphylococcal extracts with pHMB or MTSET (18), known inhibitors of the sorting reaction (11). These results suggest that sortase catalyzes the hydroxylaminolysis of LPXTG peptide in vitro.…”

Section: Staphylococcal Extracts Catalyze Hydroxylaminolysis At the Lmentioning

confidence: 80%

Purification and characterization of sortase, the transpeptidase that cleaves surface proteins of Staphylococcus aureus at the LPXTG motif

Ton‐That

Liu

Mazmanian

et al. 1999

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

530

542

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Surface proteins of Staphylococcus aureus are linked to the bacterial cell wall by sortase, an enzyme that cleaves polypeptides at the threonine of the LPXTG motif. Surface proteins can be released from staphylococci by treatment with hydroxylamine, resulting in the formation of threonine hydroxamate. Staphylococcal extracts, as well as purified sortase, catalyze the hydroxylaminolysis of peptides bearing an LPXTG motif, a reaction that can be inhibited with sulfhydryl-modifying reagents. Replacement of the single conserved cysteine at position 184 of sortase with alanine abolishes enzyme activity. Thus, sortase appears to catalyze surfaceprotein anchoring by means of a transpeptidation reaction that captures cleaved polypeptides as thioester enzyme intermediates.

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Section: Staphylococcal Extracts Catalyze Hydroxylaminolysis At the Lmentioning

confidence: 80%

Purification and characterization of sortase, the transpeptidase that cleaves surface proteins of Staphylococcus aureus at the LPXTG motif

Ton‐That

Liu

Mazmanian

et al. 1999

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

530

542

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“…We modified our experimental strategy and quenched all cellular disulfide exchange with MTSEA, a reagent that rapidly blocks cysteine thiols without attacking disulfides (32,33). When extracts of cells treated with MTSEA were analyzed by immunoblotting, DsbD C285A was observed to form an intermolecular disulfide with TrxA C35S (Fig.…”

Section: Resultsmentioning

confidence: 99%

DsbD-catalyzed Transport of Electrons across the Membrane ofEscherichia coli

Krupp

Chan

Missiakas

2001

Journal of Biological Chemistry

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Dsb proteins catalyze folding and oxidation of polypeptides in the periplasm of Escherichia coli. DsbC reduces wrongly paired disulfides by transferring electrons from its catalytic dithiol motif 98 CGYC. Genetic evidence suggests that recycling of this motif requires at least three proteins, the cytoplasmic thioredoxin reductase (TrxB) and thioredoxin (TrxA) as well as the DsbD membrane protein. We demonstrate here that electrons are transferred directly from thioredoxin to DsbD and from DsbD to DsbC. Three cysteine pairs within DsbD undergo reversible disulfide rearrangements. Our results suggest a novel mechanism for electron transport across membranes whereby electrons are transferred sequentially from cysteine pairs arranged in a thioredoxin-like motif (CXXC) to a cognate reactive disulfide.

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“…Nevertheless, SrtA ⌬N59,W194A displayed about a 2-fold reduction in the overall activity of sortase. Cleavage of the a-LPETG-d peptide still occurred in a manner requiring the active site cysteine and formation of a thiolate ion, because all hydrolysis activity of SrtA ⌬N59,W194A could be inhibited by addition of MTSET, a compound reactive with cysteine thiolate (28) (Fig. 3).…”

Section: Fig 4 Trpmentioning

confidence: 99%

Anchoring of Surface Proteins to the Cell Wall of Staphylococcus aureus

Ton‐That¹,

Mazmanian²,

Alksne³

et al. 2002

Journal of Biological Chemistry

150

105

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Surface proteins of Staphylococcus aureus are anchored to the cell wall peptidoglycan by a mechanism requiring a C-terminal sorting signal with a LPXTG motif. Sortase cleaves polypeptides between the threonine and the glycine of the LPXTG motif. The carboxyl group of threonine is subsequently amide-linked to the amino group of peptidoglycan cross-bridges. The three-dimensional structure of sortase revealed the close proximity of the catalytic site residue cysteine 184 with histidine 120; however, no structural evidence for a thiolate-imidazolium ion pair could be detected. We report that alanine substitution of either cysteine 184 or histidine 120 abolishes in vivo and in vitro sorting reactions. Further, alanine substitution of tryptophan 194, a residue that is in close proximity of histidine 120, reduces the transpeptidase activity of sortase. These results suggest a model whereby sortase forms a thiolate-imidazolium ion pair for the catalysis of its transpeptidation reaction and that the position of tryptophan 194 assists in the formation of this ion pair.

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Corrections - Simple Alkanethiol Groups for Temporary Blocking of Sulfhydryl Groups of Enzymes

Cited by 30 publications

References 0 publications

Purification and characterization of sortase, the transpeptidase that cleaves surface proteins of Staphylococcus aureus at the LPXTG motif

Purification and characterization of sortase, the transpeptidase that cleaves surface proteins of Staphylococcus aureus at the LPXTG motif

DsbD-catalyzed Transport of Electrons across the Membrane ofEscherichia coli

Anchoring of Surface Proteins to the Cell Wall of Staphylococcus aureus

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