title: Further evidence that a cell wall precursor [C(55)-MurNAc-(peptide)-GlcNAc] serves as an acceptor in a sorting reaction.
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.
authors.affiliation: Department of Infectious Disease, Wyeth-Ayerst Research, Pearl River, New York 10965, USA. ruzina@war.wyeth.com
authors.forename: Alexey
authors.initials: A
authors.lastname: Ruzin
authors.forename: Anatoly
authors.initials: A
authors.lastname: Severin
authors.forename: Frank
authors.initials: F
authors.lastname: Ritacco
authors.forename: Keiko
authors.initials: K
authors.lastname: Tabei
authors.forename: Guy
authors.initials: G
authors.lastname: Singh
authors.forename: Patricia A
authors.initials: PA
authors.lastname: Bradford
authors.forename: Marshall M
authors.initials: MM
authors.lastname: Siegel
authors.forename: Steven J
authors.initials: SJ
authors.lastname: Projan
authors.forename: David M
authors.initials: DM
authors.lastname: Shlaes
mesh_terms: D019881:Aminoacyltransferases / Q000502:physiology*; D001426:Bacterial Proteins; D002473:Cell Wall / Q000378:metabolism*; D003546:Cysteine Endopeptidases; D008565:Membrane Proteins / Q000378:metabolism; D010457:Peptidoglycan / Q000096:biosynthesis*; D013302:Streptomyces / Q000378:metabolism*; D014538:Uridine Diphosphate N-Acetylmuramic Acid / Q000031:analogs & derivatives* / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D008565:Membrane Proteins; D010457:Peptidoglycan; D014538:Uridine Diphosphate N-Acetylmuramic Acid; C069249:muramyl-NAc-(pentapeptide)pyrophosphoryl-undecaprenol; D019881:Aminoacyltransferases; C120542:sortase A; D003546:Cysteine Endopeptidases