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Antimicrobial Agents and Chemotherapy, May 2000, p. 1181-1185, Vol. 44, No. 5
0066-4804/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

A Simple Screen for Murein Transglycosylase Inhibitors

Waldemar Vollmerdagger and Joachim-Volker Höltje*

Max-Planck-Institut für Entwicklungsbiologie, Abteilung Biochemie, 72076 Tübingen, Germany

Received 27 September 1999/Returned for modification 23 December 1999/Accepted 10 February 2000

A simple assay for detection of compounds that bind to the active site in the transglycosylation domain of the essential bifunctional transglycosylase and transpeptidase penicillin-binding proteins (PBPs) is reported. The method is based on a competition with the specific transglycosylase inhibitor moenomycin. With moenomycin coupled to Affi-Gel beads, a simple filtration procedure allows the amount of labeled PBPs that bind to moenomycin beads in the presence of test substances to be determined. The PBPs can easily be labeled by the covalent binding of penicillin derivatives. Crude membrane extracts can be used as a source for the PBPs, and different kinds of labels for the penicillin-PBP complexes can be used. The assay can be adapted to high-throughput screens.

* Corresponding author. Mailing address: Max-Planck-Institut für Entwicklungsbiologie, Abteilung Biochemie, Spemannstrabeta e 35, 72076 Tübingen, Germany. Phone: 49-7071-601-412. Fax: 49-7071-601-447. E-mail: joachim-volker.hoeltje{at}tuebingen.mpg.de.

dagger Present address: Laboratory of Microbiology, The Rockefeller University, New York, NY 10021.

Antimicrobial Agents and Chemotherapy, May 2000, p. 1181-1185, Vol. 44, No. 5
0066-4804/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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