title: Screening for compounds that affect the interaction between bacterial two-component signal transduction response regulator protein and cognate promoter DNA.
abstract: Bacterial signal transduction systems can be used as drug targets. The signal transduction targets fall into two groups--sensor kinases and response regulators. Previously reported studies describe hits that were thought to inactivate sensor kinases but on closer examination were found to act elsewhere instead; a possible reason for this is that full-length sensor kinases are integral membrane proteins whose activity might reflect interaction with the cell membrane or with membrane components. We describe a model system that instead is based on the interaction between a test compound and a response regulator in a homogeneous phase reaction. In this system, response regulator-DNA complex formation and its inhibition by a test compound are measured by fluorescence polarization. The model system should be readily adaptable to drug discovery based on other bacterial two-component s transduction systems.
authors.affiliation: Department of Pharmacology, University of Wisconsin Medical School, Madison, WI, USA.
authors.forename: Matthew G
authors.initials: MG
authors.lastname: Erickson
authors.forename: Andrew T
authors.initials: AT
authors.lastname: Ulijasz
authors.forename: Bernard
authors.initials: B
authors.lastname: Weisblum
mesh_terms: D001426:Bacterial Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D004353:Drug Evaluation, Preclinical; D016984:Enterococcus faecium / Q000187:drug effects / Q000502:physiology; D005454:Fluorescence Polarization / Q000379:methods*; D010766:Phosphorylation / Q000187:drug effects; D011401:Promoter Regions, Genetic; D047428:Protein Kinase Inhibitors / Q000302:isolation & purification* / Q000494:pharmacology; D011993:Recombinant Fusion Proteins / Q000037:antagonists & inhibitors / Q000302:isolation & purification / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects*; D014157:Transcription Factors / Q000037:antagonists & inhibitors* / Q000302:isolation & purification / Q000378:metabolism; D015533:Transcriptional Activation / Q000187:drug effects*
chemical_list: D001426:Bacterial Proteins; D047428:Protein Kinase Inhibitors; D011993:Recombinant Fusion Proteins; D014157:Transcription Factors; C073902:VanR protein, bacteria; C070700:VanH protein, Enterococcus faecium