Biosynthesis of enterobacterial common antigen in Escherichia coli. In vitro synthesis of lipid-linked intermediates.
An in vitro system was developed to study the biosynthesis of enterobacterial common antigen (ECA). Membranes of Escherichia coli were found to possess an enzyme activity that catalyzes the transfer of UDP-N-acetyl-acetylglucosamine-1-phosphate from UDP-N-acetyl-glucosamine (UDP-GlcNAc) to an endogenous lipid acceptor according to the reaction UDP-GlcNAc + P-lipid----GlcNAc-PP-lipid + UMP. The lipid-linked product was tentatively identified as GlcNAc-pyrophosphorylundecaprenol (lipid I) based on a comparison of its chemical and chromatographic properties with those of authentic GlcNAc-pyrophosphorylundecaprenol. The enzyme was dependent on the presence of Mg2+ for activity, and the reaction catalyzed by the enzyme was totally inhibited by the antibiotic tunicamycin in both the forward and reverse directions. Incubation of membranes with both UDP-N-acetylmannosaminuronic acid (UDP-ManNAcA) and UDP-GlcNAc resulted in the conversion of lipid I to a more polar compound, lipid II. The synthesis of lipid II was dependent on prior synthesis of lipid I. Characterization of the saccharide moiety of lipid II resulted in the identification of this compound as ManNAcA-GlcNAc-pyrophosphorylundecaprenol.
D000942:Antigens, Bacterial / Q000096:biosynthesis*; D004926:Escherichia coli / Q000276:immunology*; D006017:Glycolipids / Q000096:biosynthesis; D008055:Lipids / Q000096:biosynthesis*; D010770:Phosphotransferases / Q000378:metabolism; D017855:Transferases (Other Substituted Phosphate Groups); D014415:Tunicamycin / Q000494:pharmacology; D014537:Uridine Diphosphate N-Acetylglucosamine / Q000378:metabolism; D014539:Uridine Diphosphate Sugars / Q000378:metabolism
D000942:Antigens, Bacterial; D006017:Glycolipids; D008055:Lipids; D014539:Uridine Diphosphate Sugars; C017286:enterobacterial common antigen; C052604:N-acetylglucosamine-pyrophosphorylundecaprenol; D014415:Tunicamycin; C020949:uridine diphosphate N-acetyl-D-mannosaminuronic acid; D014537:Uridine Diphosphate N-Acetylglucosamine; C052605:N-acetylmannosaminuronic acid-N-acetylglucosamine-pyrophosphorylundecaprenol; D010770:Phosphotransferases; D017855:Transferases (Other Substituted Phosphate Groups); C023222:UDPacetylglucosamine-dolichyl-phosphate acetylglucosamine-1-phosphate transferase