The Peptide Subunit N <sup>α</sup>-(L-Alanyl-D-isoglutaminyl)-L-lysyl -D-alanine in Cell Wall Peptidoglycans of Staphylococcus aureus Strain Copenhagen, Micrococcus roseus R 27, and Sreptococcus pyogenes Group A, Type 14

Ghuysen, Jean‐Marie; Muñoz, Emilio; Leyh‐Bouille, Mélina; Petit, Jean‐François; Heymann, H; Bricas, E.; Lefrancier, P.

doi:10.1021/bi00876a004

Cited by 68 publications

(43 citation statements)

References 26 publications

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“…2) reacted weakly with anti-N-acetylglucosamine antibody. These findings suggest the presence of a region of the streptococcal cell wall with a high degree of cross-linking (25,26) that is resistant to muralytic enzyme, thus trapping a portion of the M molecule within this resistant core. In Bacillus subtilis, an enzyme-resistant fraction of the cell wall has been described (6) and attributed to the cell wall ends, which are older and thus are likely to be more cross-linked.…”

Section: Discussionmentioning

confidence: 91%

“…Evidence peptidoglycan is L-Ala-L-Ala (13), the absence of additional alanines from the amino acid composition of the wall-associated fragment (Table 1), which would be contributed by the peptide portion of the peptidoglycan, rules out any linkage through this moiety. Thus, although previous studies suggested a covalent interaction between the M protein and the cell wall (11), it is likely that the C-terminal region of the M protein is not covalently linked to but is intercalated within the highly cross-linked peptidoglycan (25). The nearly regular placement of prolines and glycines within this region suggests that some sort of specific structure may be necessary for this interaction (12).…”

Section: Discussionmentioning

confidence: 96%

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Isolation and characterization of the cell-associated region of group A streptococcal M6 protein

1988

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DNA sequence analysis of the complete M6 protein gene revealed 19 hydrophobic amino acids at the C terminus which could act as a membrane anchor and an adjacent proline-and glycine-rich region likely to be located in the cell wall. To define this region within the cell wall and its role in attaching the molecule to the cell, we isolated the cell-associated fragment of the M protein. Assuming that the cell-associated region of the M protein would be embedded within the' wall and thus protected from trypsin digestion, cells were digested with this enzyme, and the wall-associated M protein fragmnent was released by phage lysin digestion of the peptidoglycan. With antibody probes prepared to synthetic peptides of C-terminal sequences, a cell wallassociated M protein fragment (molecular weight, 16,000) was identified and purified. Amino acid sequence analysis placed the N terminus of the 16,000-molecular-weight fragment at residue 298 within the M sequence. Amino acid composition of this peptide was consistent with a C-terminal sequence lacking the membrane anchor. Antibody studies of nitrous acid-extracted whole bacteria suggested that, in addition to the peptidoglycan-associated region, a 65-residue helical segment of the C-terminal domain of the M protein is embedded within the carbohydrate moiety of the cell wall. Since no detectable amino sugars were associated with the wall-associated fragment, the C-terminal region of the M6 molecule is likely to be intercalated within the cross-linked peptidoglycan and not covalently linked to it. Because the C-terminal region of the M molecule is highly homologous to the C-terminal end of protein A from staphylococci and protein G from streptococci, it is likely that the mechanism of attachment of these proteins to the cell wall is conserved.M protein is an antiphagocytic molecule located on the surface of the group A streptococcal cell wall (21,31). From physicochemical and amino acid sequence analyses of the M5 (22), M6 (12, 16), and M24 proteins (2), it is clear that M protein is composed of two alpha-helical protein chains wound around each other to form a coiled-coil structure (12,27). Although the majority of structural and immunochemical data on the M protein is derived from an amino-terminal fragment released from the streptococcus by limited pepsin digestion (2,3,(22)(23)(24)27), information regarding the Cterminal portion of this molecule is beginning to emerge (12,16,17,19,20,28).DNA sequence analysis of the M6 gene revealed the complete amino acid sequence of this M molecule, which identified three different repeat regions (16). In addition, it revealed a region at the C-terminal end which is likely to be responsible for attaching the M molecule to the cell. This segment of the M protein contains a potential membrane anchor region composed of 19 hydrophobic amino acids with a charged tail probably extending into the cytoplasm. Adjacent to the membrane anchor is a proline-and glycine-rich region which is likely to be located within the cell wall. The nearly regula...

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Section: Discussionmentioning

confidence: 91%

Section: Discussionmentioning

confidence: 96%

Isolation and characterization of the cell-associated region of group A streptococcal M6 protein

1988

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“…Penicillin binding proteins polymerize peptidoglycan precursors into linear glycan strands that are cross-linked with neighboring strands, thereby generating a single large murein macromolecule (23,46,53). Staphylococcus aureus peptidoglycan is composed of N-acetylmuramoyl-(L-Ala-D-iGln-L-Lys(Gly 5 )-D-Ala)-(␤1-4)-N-acetylglucosamine (7,13,14,32). Polymerization of repeating disaccharide units, N-acetylmuramoyl-(␤1-4)-N-acetylglucosamine (MurNAc-GlcNAc), generates the glycan strands (3,8).…”

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confidence: 99%

Distribution of Protein A on the Surface ofStaphylococcus aureus

2007

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Surface proteins of Staphylococcus aureus fulfill many important roles during the pathogenesis of human infections and are anchored to the cell wall envelope by sortases. Although the chemical linkage of proteins to cell wall cross bridges is known, the mechanisms whereby polypeptides are distributed on the staphylococcal surface have not been revealed. We show here that protein A, the ligand of immunoglobulin, is unevenly distributed over the staphylococcal surface. Upon removal with trypsin, newly synthesized polypeptide is deposited at two to four discrete foci. During subsequent growth, protein A appears to be slowly distributed from these sites. When viewed through multiple focal planes by laser scanning microscopy, protein A foci are arranged in a circle surrounding the bacterial cell. This pattern of distribution requires the LPXTG sorting signal of protein A as well as sortase A, the transpeptidase that anchors polypeptides to cell wall cross bridges. A model is presented whereby protein A deposition at discrete sites coupled with cell wall synthesis enables distribution of protein A on the staphylococcal surface.

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“…Penicillin-binding proteins (PBPs), another group of bacterial transpeptidases, cleave cell-wall peptides at the amide bond of D-Ala-D-Ala (16,17). The carboxyl group of the cleaved peptide is ester-linked to the hydroxyl group of the active-site serine residue (18,19).…”

mentioning

confidence: 99%

Structure of sortase, the transpeptidase that anchors proteins to the cell wall of Staphylococcus aureus

Ilangovan

Ton‐That

Iwahara

et al. 2001

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

285

366

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Surface proteins of Gram-positive bacteria play important roles during the pathogenesis of human infections and require sortase for anchoring to the cell-wall envelope. Sortase cleaves surface proteins at the LPXTG motif and catalyzes the formation of an amide bond between the carboxyl group of threonine (T) and the amino group of cell-wall crossbridges. The NMR structure of sortase reveals a unique ␤-barrel structure, in which the active-site sulfhydryl of cysteine-184 is poised for ionization by histidine-120, presumably enabling the resultant thiolate to attack the LPXTG peptide. Calcium binding near the active site stimulates catalysis, possibly by altering the conformation of a surface loop that recognizes newly translocated polypeptides. The structure suggests a mechanistic relationship to the papain͞cathepsin proteases and should facilitate the design of new antiinfective agents.

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The Peptide Subunit N ^α-(L-Alanyl-D-isoglutaminyl)-L-lysyl -D-alanine in Cell Wall Peptidoglycans of Staphylococcus aureus Strain Copenhagen, Micrococcus roseus R 27, and Sreptococcus pyogenes Group A, Type 14

Cited by 68 publications

References 26 publications

Isolation and characterization of the cell-associated region of group A streptococcal M6 protein

Isolation and characterization of the cell-associated region of group A streptococcal M6 protein

Distribution of Protein A on the Surface ofStaphylococcus aureus

Structure of sortase, the transpeptidase that anchors proteins to the cell wall of Staphylococcus aureus

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The Peptide Subunit N α-(L-Alanyl-D-isoglutaminyl)-L-lysyl -D-alanine in Cell Wall Peptidoglycans of Staphylococcus aureus Strain Copenhagen, Micrococcus roseus R 27, and Sreptococcus pyogenes Group A, Type 14

Cited by 68 publications

References 26 publications

Isolation and characterization of the cell-associated region of group A streptococcal M6 protein

Isolation and characterization of the cell-associated region of group A streptococcal M6 protein

Distribution of Protein A on the Surface ofStaphylococcus aureus

Structure of sortase, the transpeptidase that anchors proteins to the cell wall of Staphylococcus aureus

Contact Info

Product

Resources

About

The Peptide Subunit N ^α-(L-Alanyl-D-isoglutaminyl)-L-lysyl -D-alanine in Cell Wall Peptidoglycans of Staphylococcus aureus Strain Copenhagen, Micrococcus roseus R 27, and Sreptococcus pyogenes Group A, Type 14