Perry, Adrienne M. and Ton-That, Hung and Mazmanian, Sarkis K. and Schneewind, Olaf (2002) Anchoring of Surface Proteins to the Cell Wall of Staphylococcus aureus. III. Lipid II is an in vivo peptidoglycan substrate for sortase-catalyzed surface protein anchoring. Journal of Biological Chemistry, 277 (18). pp. 16241-16248. ISSN 0021-9258. http://resolver.caltech.edu/CaltechAUTHORS:PERjbc02
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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 an LPXTG motif. Surface proteins are first synthesized in the bacterial cytoplasm and then transported across the cytoplasmic membrane. Cleavage of the N-terminal signal peptide of the cytoplasmic surface protein P1 precursor generates the extracellular P2 species, which is the substrate for the cell wall anchoring reaction. Sortase, a membrane-anchored transpeptidase, cleaves P2 between the threonine (T) and the glycine (G) of the LPXTG motif and catalyzes the formation of an amide bond between the carboxyl group of threonine and the amino group of cell wall cross-bridges. We have used metabolic labeling of staphylococcal cultures with [32P]phosphoric acid to reveal a P3 intermediate. The 32P-label of immunoprecipitated surface protein is removed by treatment with lysostaphin, a glycyl-glycine endopeptidase that separates the cell wall anchor structure. Furthermore, the appearance of P3 is prevented in the absence of sortase or by the inhibition of cell wall synthesis. 32P-Labeled cell wall anchor species bind to nisin, an antibiotic that is known to form a complex with lipid II. Thus, it appears that the P3 intermediate represents surface protein linked to the lipid II peptidoglycan precursor. The data support a model whereby lipid II-linked polypeptides are incorporated into the growing peptidoglycan via the transpeptidation and transglycosylation reactions of cell wall synthesis, generating mature cell wall-linked surface protein.
|Additional Information:||Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc. Received for publication, September 24, 2001, and in revised form, February 18, 2002. Originally published In Press as doi:10.1074/jbc.M109194200 on February 20, 2002 This work was supported by United States Public Health Service Grant AI38897. This is Paper III in the series Anchoring of Surface Proteins to the Cell Wall of Staphylococcus aureus. Refs. 51 and 52 are Papers I and II, respectively.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.|
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|Deposited On:||15 Nov 2006|
|Last Modified:||26 Dec 2012 09:17|
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