Further evidence that a cell wall precursor [C(55)-MurNAc-(peptide)-GlcNAc] serves as an acceptor in a sorting reaction.
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.
Ruzin
Alexey
A
Department of Infectious Disease, Wyeth-Ayerst Research, Pearl River, New York 10965, USA. ruzina@war.wyeth.com
Severin
Anatoly
A
Ritacco
Frank
F
Tabei
Keiko
K
Singh
Guy
G
Bradford
Patricia A
PA
Siegel
Marshall M
MM
Projan
Steven J
SJ
Shlaes
David M
DM
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*
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