Anchor Structure of Staphylococcal Surface Proteins

Ton‐That, Hung; Faull, Kym F.; Schneewind, Olaf

doi:10.1074/jbc.272.35.22285

Cited by 123 publications

(68 citation statements)

References 51 publications

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“…Separation was carried out with a gradient from 1% to 41% CH 3 CN (0.1% trifluoroacetic acid) in 41 min and from 41% to 100% for 10 min at a flow rate of 0.3 ml͞min. Elution of peptides was monitored at 206 and 475 nm and fractions were collected every minute, dried under vacuum, and stored at 4°C for electrospray ionization-MS analysis, as described (6,28).…”

Section: Characterization Of Cleaved Productsmentioning

confidence: 99%

“…During cell-wall anchoring, surface proteins of Staphylococcus aureus are cleaved between the threonine and the glycine of the LPXTG motif (4). The carboxyl group of threonine is amide-linked to the amino group of the pentaglycine crossbridges, thereby anchoring the C-terminal end of surface proteins to the staphylococcal cell-wall peptidoglycan (5,6). It seems that lipid II [undecaprenyl pyrophosphate-MurNAc(-L-Ala-D-iGln-L-Lys(NH 2 -Gly 5 )-D-Ala-D-Ala)-␤1-4-GlcNAc)], a membrane-anchored precursor of cell-wall synthesis (7,8), serves as a substrate for surface protein anchoring (9).…”

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confidence: 99%

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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.

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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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Section: Characterization Of Cleaved Productsmentioning

confidence: 99%

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.

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281

366

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“…Sortase A, a transpeptidase with an active site cysteine, cleaves the sorting signal between the threonine and the glycine of the LPXTG motif (4 -7). Sortase catalyzes the formation of an amide bond between the carboxyl group of threonine and the amino group of the cell wall cross-bridge, a pentaglycyl moiety in staphylococci (8,9). Several observations suggest that lipid II, a membrane anchored intermediate of cell wall synthesis, functions as the peptidoglycan substrate of sortase A (10,11).…”

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confidence: 99%

Crystal Structures of Staphylococcus aureus Sortase A and Its Substrate Complex

Zong

Bice

Ton‐That

et al. 2004

Journal of Biological Chemistry

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“…Sortase A cuts surface protein precursors at their LPXTG sorting signal and accepts lipid II as a nucleophile, thereby incorporating proteins into the peptidoglycan synthesis pathway (15,34,35). Sortase B recognizes surface protein precursors with distinct sorting signals (NPQTN motif in S. aureus) and employs cross-linked cell wall as a nucleophile (26,36).…”

Section: Discussionmentioning

confidence: 99%

Cell Wall Anchor Structure of BcpA Pili in Bacillus anthracis

Budzik¹,

Schneewind

2008

Journal of Biological Chemistry

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Assembly of pili in Gram-positive bacteria and their attachment to the cell wall envelope are mediated by sortases. In Bacillus cereus and its close relative Bacillus anthracis, the major pilin protein BcpA is cleaved between the threonine and the glycine of its C-terminal LPXTG motif sorting signal by the pilin-specific sortase D. The resulting acyl enzyme intermediate is relieved by the nucleophilic attack of the side-chain amino group of lysine within the YPKN motif of another BcpA subunit. Cell wall anchoring of assembled BcpA pili requires sortase A, which also cleaves the LPXTG sorting signal of BcpA between its threonine and glycine residues. We show here that sortases A and D require only the C-terminal sorting signal of BcpA for substrate cleavage. Unlike sortase D, which accepts the YPKN motif as a nucleophile, sortase A forms an amide bond between the BcpA C-terminal carboxyl group of threonine and the sidechain amino group of diaminopimelic acid within the cell wall peptidoglycan of bacilli. These results represent the first demonstration of a cell wall anchor structure for pili, which are deposited by sortase A into the envelope of many different microbes.Many Gram-positive bacteria use pili, protein fibers that project from microbial surfaces, to adhere to and invade host tissues or form biofilms (1). Gram-positive pili are assembled from pilin subunits that are synthesized as precursors with N-terminal signal peptides and C-terminal sorting signals in the bacterial cytoplasm (P1) (2). The signal peptide is cleaved and precursors translocated through the bacterial membrane by the Sec machinery, thereby generating the P2 precursor (3, 4). Assembly of pili involves the polymerization of the major pilin protein, a reaction that is catalyzed by pilinspecific sortase (2, 5), designated sortase D in pathogenic Bacillus sp. (6). Pilin-specific sortases perform a transpeptidation reaction, whereby the C-terminal LPXTG motif sorting signal of the major pilin protein, BcpA in bacilli, is cleaved between the threonine and the glycine residue (4). Intermediary product of this reaction is a thioester-linked acyl-enzyme between the active site cysteine residue of sortase and the carboxyl group of threonine (7,8). Nucleophilic attack of the ⑀-amino group of lysine within the YPKN pilin motif of another BcpA subunit resolves the sortase D intermediate and generates the amide bond between the C-terminal carboxyl group of threonine and the side-chain amino group of lysine of adjacent BcpA subunits (4). Conservation of pilin-specific sortase, the YPKN motif, and C-terminal sorting signal in the major subunit of pilin proteins suggests that pilus assembly occurs by a universal mechanism in Gram-positive bacteria (2).Pili are anchored to the cell wall envelope of Gram-positive bacteria by a mechanism that requires sortase A (6, 9 -11). Sortase A is otherwise known to cut the C-terminal LPXTG sorting signal of surface proteins and immobilize anchored products in the cell wall envelope of 13). In Staphylococcus aureus, s...

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Anchor Structure of Staphylococcal Surface Proteins

Cited by 123 publications

References 51 publications

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

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

Crystal Structures of Staphylococcus aureus Sortase A and Its Substrate Complex

Cell Wall Anchor Structure of BcpA Pili in Bacillus anthracis

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