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A Microtiter Plate-Based Assay for Inhibitors of Penicillin-Binding Protein 2a from Methicillin Resistant Staphylococcus aureus.

Antimicrob Agents Chemother.. 2011-06; 
Sudheer Bobba, V. K. Chaithanya Ponnaluri, Mridul Mukherji, and William G. Gutheil. Division of Pharmaceutical Sciences, University of Missouri-Kansas City, 2464 Charlotte Street, Kansas City, Missouri 64108, USA.
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摘要

Penicillin-binding protein 2a (PBP2a), the molecular determinant for high-level β-lactam resistance in methicillin-resistant Staphylococcus aureus (MRSA), is intrinsically resistant to most β-lactam antibiotics. The development and characterization of new inhibitors targeting PBP2a would benefit from an effective and convenient assay for inhibitor binding. This study was directed toward the development of a fluorescently detected β-lactam binding assay for PBP2a from MRSA. Biotinylated ampicillin and biotinylated cephalexin were tested as tagging reagents for fluorescence detection by using a streptavidin-horseradish peroxidase conjugate. Both bound surprisingly well to PBP2a, with binding consta... More

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