Escherichia coli murein transglycosylase. Purification by affinity chromatography and interaction with polynucleotides.
Escherichia coli murein transglycosylase, a potential autolysin which splits the sugar chains of the murein sacculus, was rapidly purified from a crude cell extract by sequential chromatography on columns of blue Sepharose and poly(U)-Sepharose. In accordance with the binding to blue Sepharose and poly(U)-Sepharose, the transglycosylase is inhibited by Cibacron blue F3G-A, the affinity ligand of blue Sepharose, and also by polynucleotides, the latter, however, with varying efficiency. Among the polynucleotides tested, single-stranded DNA was found to be one of the most potent inhibitors. When bound to a blue Sepharose column, transglycosylase could be displaced from the column with single-stranded DNA. Taken together, these results point to a polynucleotide binding area on the transglycosylase molecule. Some aspects of the blue Sepharose affinity chromatography and the possible biological significance of the transglycosylase are discussed.
D002846:Chromatography, Affinity; D004926:Escherichia coli / Q000201:enzymology*; D016695:Glycosyltransferases; D007700:Kinetics; D008970:Molecular Weight; D011119:Polynucleotides / Q000494:pharmacology; D013329:Structure-Activity Relationship; D014166:Transferases / Q000302:isolation & purification* / Q000378:metabolism
D011119:Polynucleotides; D014166:Transferases; D016695:Glycosyltransferases; C021815:murein transglycosylase