title: Use of a semisynthetic epitope to probe histidine kinase activity and regulation.
abstract: Histidine-aspartic acid phosphotransfer pathways are central components of prokaryotic signal transduction pathways and are also found in many eukaryotes. Tools to study histidine kinases, however, are currently quite limited. In this article, we present a new tool to study histidine-aspartic acid phosphotransfer pathways. We show that many histidine kinases will accept ATPgammaS as a substrate to form a stable thiophosphohistidine even when they do not form stable phosphohistidines using the natural substrate ATP. An antibody that has previously been used to detect thiophosphorylated serine, threonine, and tyrosine residues is shown to recognize thiophosphohistidine and thiophosphoaspartic acid residues. Histidine kinase autothiophosphorylation is regulated by other protein sensor domains in the same way as autophosphorylation, and thiophosphate is transferred to downstream aspartic acid containing response regulators.
authors.affiliation: Department of Chemistry, University of California, Berkeley, Berkeley, CA 94720, USA.
authors.forename: Hans K
authors.initials: HK
authors.lastname: Carlson
authors.forename: Lars
authors.initials: L
authors.lastname: Plate
authors.forename: Mark S
authors.initials: MS
authors.lastname: Price
authors.forename: Jasmina J
authors.initials: JJ
authors.lastname: Allen
authors.forename: Kevan M
authors.initials: KM
authors.lastname: Shokat
authors.forename: Michael A
authors.initials: MA
authors.lastname: Marletta
mesh_terms: D000255:Adenosine Triphosphate / Q000031:analogs & derivatives* / Q000378:metabolism; D003001:Cloning, Molecular; D000939:Epitopes / Q000737:chemistry*; D004926:Escherichia coli / Q000201:enzymology; D006639:Histidine / Q000031:analogs & derivatives / Q000032:analysis / Q000276:immunology; D000071677:Histidine Kinase; D011494:Protein Kinases / Q000032:analysis* / Q000378:metabolism*
chemical_list: D000939:Epitopes; C022571:adenosine 5'-O-(3-thiotriphosphate); D006639:Histidine; D000255:Adenosine Triphosphate; D011494:Protein Kinases; D000071677:Histidine Kinase