Isolation by covalent affinity chromatography of the penicillin-binding components from membranes of Bacillus subtilis.
An affinity chromatography technique was developed to isolate the five penicillin-binding components present in Bacillus subtilis membranes. The proteins were solubilized by the detergent Nonidet P-40, bound covalently to penicillin-substituted Sepharose, and subsequently eluted from the matrix with neutral hydroxylamine, which cleaves the penicilloyl-enzyme bond. Penicillin binding-component V, the D-alanine carboxypeptidase, makes up 1% of the total membrane protein. A modification of the above procedure enabled this enzyme to be obtained from the membrane in pure form in a single step with 50% overall recovery of enzymatic activity.
D000409:Alanine; D001412:Bacillus subtilis / Q000032:analysis* / Q000201:enzymology; D001426:Bacterial Proteins / Q000302:isolation & purification* / Q000378:metabolism; D001665:Binding Sites; D002247:Carbon Isotopes; D002268:Carboxypeptidases / Q000302:isolation & purification; D002462:Cell Membrane / Q000032:analysis; D002845:Chromatography; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D010400:Penicillin G / Q000378:metabolism*; D011485:Protein Binding
D001426:Bacterial Proteins; D002247:Carbon Isotopes; D002268:Carboxypeptidases; D000409:Alanine; D010400:Penicillin G