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Journal of Biomolecular Screening, Vol. 10, No. 3, 270-274 (2005)
DOI: 10.1177/1087057104273930
© 2005 Society for Biomolecular Sciences

Bacterial 2-Component Signal Transduction Systems: A Fluorescence Polarization Screen for Response Regulator-Protein Binding

Matthew G. Erickson

Andrew T. Ulijasz

Bernard Weisblum

Pharmacology Department, University of Wisconsin Medical School, Madison

Two-component signal transduction systems are the primary means by which bacteria sense environmental change and integrate an adaptive response. In pathogenic bacteria, 2-component signal transduction (TCST) kinases are involved in the expression of virulence and antibiotic resistance. This makes bacterial TCST systems attractive targets for pharmacologic intervention. This paper describes a fluorescence polarization assay that quantifies the binding between bacterial DNA promoter segments and their cognate response regulator proteins. Using the Van RSTCST system from Enterococcus faecium, which encodes vancomycin resistance, the authors demonstrate inhibition of response regulator protein/promoter segment binding with a known inhibitor. Observed binding constants were comparable to those reported in surface plasmon resonance measurements and gel shift measurements.

Key Words: fluorescence polarization • response regulator • bacterial signal transduction systems • antibiotic resistance


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