Work

A High-Throughput TNP-ATP Displacement Assay for Screening Inhibitors of ATP-Binding in Bacterial Histidine Kinases

Year: 2010

Type: article

Abstract: Bacterial histidine kinases (HK) are members of the GHKL superfamily, which share a unique adenosine triphosphate (ATP)-binding Bergerat fold. Our previous studies have shown that Gyrase, Hsp90, MutL (GHL) inhibitors bind to the ATP-binding pocket of HK and may provide lead compounds for the design of novel antibiotics targeting these kinases. In this article, we developed a competition assay using the fluorescent ATP analog, 2',3'-O-(2,4,6-trinitrophenyl) adenosine 5'-triphosphate. The method can be used for high-throughput screening of compound libraries targeting HKs or other ATP-binding proteins. We utilized the assay to screen a library of GHL inhibitors targeting the bacterial HK PhoQ, and discuss the applications of the 2',3'-O-(2,4,6-trinitrophenyl) adenosine 5'-triphosphate competition assay beyond GHKL inhibitor screening.

Source: Assay and Drug Development Technologies

DOI: 10.1089/adt.2010.0289

Authors: Michael T. Guarnieri, Brian S. J. Blagg, Rui Zhao

Institutions: University of Colorado Denver, University of Kansas

Language: English

Topic: Bacterial Genetics and Biotechnology

Subfield: Genetics

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

Open Access status: green