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Chemical Probes For Histidine Kinase Profiling And Inhibitor Discovery
Author(s) -
Carlson Erin
Publication year - 2016
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.30.1_supplement.100.1
Subject(s) - autophosphorylation , kinase , kinome , phosphorylation , biochemistry , small molecule , signal transduction , microbiology and biotechnology , chemistry , biology , chemical biology , drug discovery , protein kinase a
Adaptive bacterial signaling is largely regulated by two‐component systems (TCSs). Comprised of histidine kinases (HKs) and response regulators (RRs), these phosphorylation cascades result in myriad intracellular responses that include various pathogenic mechanisms. Despite their abundance in Gram‐negative, Gram‐positive, and Mycobacterial organisms, little is known about global TCS regulation. We are working to address this deficiency by generation of probes to profile HK autophosphorylation activity. Significantly, we have identified the first non‐radioactive activity‐based probe for HK signaling, BODIPY‐FL‐ATP‐gamma‐S, a fluorescent adenosine triphosphate (ATP)‐based probe that exploits thiophosphorylation to afford greater stability and thus more facile detection of protein activation. This and other ATP‐gamma‐S‐based probes are in development for the profiling of HK activity from bacterial proteomes. We also seek to inhibit TCS signaling networks and thereby severely attenuate bacteria's ability to cause infection. We have targeted the highly conserved ATP‐binding domain present across the HK superfamily. An ADP‐based probe enabled us to pursue new inhibitor scaffolds through the execution of a high‐throughput screen of ~75,000 diverse small molecules and natural product extracts. As a result, nine lead compounds have been identified for deactivation of multiple HKs. Scaffolds identified in these studies contribute to the foundation for HK inhibitor development, supporting a new class of antibiotics with a novel mechanism of action. Together, our chemical probes will continue to advance our current understanding of global HK regulation and inhibition.