Further evidence that a cell wall precursor [C(55)-MurNAc-(peptide)-GlcNAc] serves as an acceptor in a sorting reaction |
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Authors: | Ruzin Alexey Severin Anatoly Ritacco Frank Tabei Keiko Singh Guy Bradford Patricia A Siegel Marshall M Projan Steven J Shlaes David M |
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Institution: | Department of Infectious Disease, Wyeth-Ayerst Research, Pearl River, New York 10965, USA. ruzina@war.wyeth.com |
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Abstract: | Previous studies suggested that a Gly-containing branch of cell wall precursor C(55)-MurNAc-(peptide)-GlcNAc], which is often referred to as lipid II, might serve as a nucleophilic acceptor in sortase-catalyzed anchoring of surface proteins in Staphylococcus aureus. To test this hypothesis, we first simplified the procedure for in vitro biosynthesis of Gly-containing lipid II by using branched UDP-MurNAc-hexapeptide isolated from the cytoplasm of Streptomyces spp. Second, we designed a thin-layer chromatography-based assay in which the mobility of branched but not linear lipid II is shifted in the presence of both sortase and LPSTG-containing peptide. These results and those of additional experiments presented in this study further suggest that lipid II indeed serves as a natural substrate in a sorting reaction. |
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