title: Characterization of Wall Teichoic Acid Degradation by the Bacteriophage ϕ29 Appendage Protein GP12 Using Synthetic Substrate Analogs.
abstract: The genetics and enzymology of the biosynthesis of wall teichoic acid have been the extensively studied, however, comparatively little is known regarding the enzymatic degradation of this biological polymer. The GP12 protein from the Bacillus subtilis bacteriophage ϕ29 has been implicated as a wall teichoic acid hydrolase. We have studied the wall teichoic acid hydrolase activity of pure, recombinant GP12 using chemically defined wall teichoic acid analogs. The GP12 protein had potent wall teichoic acid hydrolytic activity in vitro and demonstrated ∼13-fold kinetic preference for glycosylated poly(glycerol phosphate) teichoic acid compared with non-glycosylated. Product distribution patterns suggested that the degradation of glycosylated polymers proceeded from the hydroxyl terminus of the polymer, whereas hydrolysis occurred at random sites in the non-glycosylated polymer. In addition, we present evidence that the GP12 protein possesses both phosphodiesterase and phosphomonoesterase activities.
authors.affiliation: From the Department of Biochemistry and Biomedical Sciences, Michael G. DeGroote Institute for Infectious Disease Research, McMaster University, Hamilton, Ontario L8N 3Z5, Canada and.
authors.forename: Cullen L
authors.initials: CL
authors.lastname: Myers
authors.affiliation: From the Department of Biochemistry and Biomedical Sciences, Michael G. DeGroote Institute for Infectious Disease Research, McMaster University, Hamilton, Ontario L8N 3Z5, Canada and.
authors.forename: Ronald G
authors.initials: RG
authors.lastname: Ireland
authors.affiliation: the Department of Chemistry, Vassar College, Poughkeepsie, New York 12604.
authors.forename: Teresa A
authors.initials: TA
authors.lastname: Garrett
authors.affiliation: From the Department of Biochemistry and Biomedical Sciences, Michael G. DeGroote Institute for Infectious Disease Research, McMaster University, Hamilton, Ontario L8N 3Z5, Canada and ebrown@mcmaster.ca.
authors.forename: Eric D
authors.initials: ED
authors.lastname: Brown
mesh_terms: D017103:Bacillus Phages / Q000201:enzymology; D001412:Bacillus subtilis / Q000737:chemistry / Q000821:virology; D055162:Biocatalysis; D002473:Cell Wall / Q000737:chemistry; D007700:Kinetics; D010744:Phosphoric Monoester Hydrolases / Q000737:chemistry*; D013682:Teichoic Acids / Q000737:chemistry*; D014764:Viral Proteins / Q000737:chemistry*
chemical_list: D013682:Teichoic Acids; D014764:Viral Proteins; D010744:Phosphoric Monoester Hydrolases
keywords: Bacillus subtilis; bacteriophage; enzyme; enzyme mechanism; enzyme turnover; phosphodiesterases; wall teichoic acid