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Journal of Bacteriology, April 2002, p. 2141-2147, Vol. 184, No. 8
0021-9193/02/$04.00+0 DOI: 10.1128/JB.184.8.2141-2147.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.
Further Evidence that a Cell Wall Precursor [C55-MurNAc-(Peptide)-GlcNAc] Serves as an Acceptor in a Sorting Reaction
Alexey Ruzin,* Anatoly Severin, Frank Ritacco, Keiko Tabei, Guy Singh, Patricia A. Bradford, Marshall M. Siegel, Steven J. Projan, and David M. Shlaes
Wyeth-Ayerst Research, Pearl River, New York 10965
Received 17 September 2001/
Accepted 11 January 2002
Previous studies suggested that a Gly-containing branch of cell wall precursor [C55-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.
* Corresponding author. Mailing address: Department of Infectious Disease, Wyeth-Ayerst Research, 401 North Middletown Rd., Pearl River, NY 10965. Phone: (845) 732-5472. Fax: (845) 732-5671. E-mail:
ruzina{at}war.wyeth.com.
Journal of Bacteriology, April 2002, p. 2141-2147, Vol. 184, No. 8
0021-9193/02/$04.00+0 DOI: 10.1128/JB.184.8.2141-2147.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.
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