PMID:10930738
| Citation |
von Rechenberg, M and Höltje, JV (2000) Two-step procedure for purification and separation of the essential penicillin-binding proteins PBP 1A and 1Bs of Escherichia coli. FEMS Microbiol. Lett. 189:201-4 |
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| Abstract |
The penicillin-binding proteins PBP 1A and 1Bs are the essential murein polymerases of Escherichia coli. Purification of these membrane-bound bifunctional transglycosylase-transpeptidases was a major obstacle in studying the details of both enzymatic reactions. Here we describe a simple, highly specific affinity chromatography method that takes advantage of the availability of the specific inhibitor of the transglycosylase site moenomycin A in order to enrich PBP 1A and 1Bs in one step from crude membrane preparations. Separation of PBP 1A from PBP 1Bs is achieved in a second step employing cation exchange chromatography yielding enzymatically active native murein polymerases. |
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| Keywords |
Bacterial Proteins; Carrier Proteins; Escherichia coli/metabolism; Hexosyltransferases/isolation & purification; Hexosyltransferases/metabolism; Multienzyme Complexes/isolation & purification; Multienzyme Complexes/metabolism; Muramoylpentapeptide Carboxypeptidase; Penicillin-Binding Proteins; Peptidyl Transferases/isolation & purification; Peptidyl Transferases/metabolism |
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