title: Purification and in vitro phosphorylation of HupT, a regulatory protein controlling hydrogenase gene expression in Rhodobacter capsulatus.
abstract: The HupT protein of Rhodobacter capsulatus, involved in negative regulation of hydrogenase gene expression, is predicted to be a histidine kinase on the basis of sequence comparisons. The protein was overproduced in Escherichia coli, purified to homogeneity, and demonstrated to autophosphorylate in vitro in the presence of [gamma-32P]ATP. An H217N hupt mutant was constructed, and the mutant protein was shown to have lost kinase activity. This result, and the fact that the phosphoryl group in phosphorylated HupT appeared to be bound to an N atom, support the suggestion from sequence comparisons that HupT is a histidine kinase, which can autophosphorylate on the His217 residue.
authors.affiliation: CEA/Grenoble, Département de Biologie Moléculaire et Structurale, France.
authors.forename: S
authors.initials: S
authors.lastname: Elsen
authors.forename: A
authors.initials: A
authors.lastname: Colbeau
authors.forename: P M
authors.initials: PM
authors.lastname: Vignais
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics; D015964:Gene Expression Regulation, Bacterial; D006864:Hydrogenase / Q000096:biosynthesis; D010766:Phosphorylation; D011494:Protein Kinases / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism; D016354:Rhodobacter capsulatus / Q000201:enzymology* / Q000235:genetics
chemical_list: D001426:Bacterial Proteins; C084201:HupT protein, Rhodobacter capsulatus; D011994:Recombinant Proteins; D006864:Hydrogenase; D011494:Protein Kinases