title: Substrate specificity and identification of functional groups of homoserine kinase from Escherichia coli.
abstract: Homoserine kinase, an enzyme in the aspartate pathway of amino acid biosynthesis in Escherichia coli, catalyzes the conversion of L-homoserine to L-homoserine phosphate. This enzyme has been found to have broad substrate specificity, including the phosphorylation of L-homoserine analogs where the carboxyl functional group at the alpha-position has been replaced by an ester or by a hydroxymethyl group. Previous pH profile studies [Huo. X., & Viola, R. E. (1996) Arch. Biochem. Biophys. 330, 373-379] and chemical modification studies have suggested the involvement of histidinyl, lysyl, and argininyl residues in the catalytic activity of the enzyme. With the assistance of sequence alignments, several potential amino acids have been targeted for examination. Site-directed mutagenesis studies have confirmed a role for arginine-234 in the binding of the carboxyl group of L-homoserine, and the involvement of two histidine at the homoserine binding site. Mutations at these sites have led to the decoupling of the kinase activity from an inherent ATPase activity in the enzyme, and suggest the presence of independent domains for the binding of each substrate in homoserine kinase.
authors.affiliation: Department of Chemistry, University of Akron, Ohio 44325-3601, USA.
authors.forename: X
authors.initials: X
authors.lastname: Huo
authors.forename: R E
authors.initials: RE
authors.lastname: Viola
mesh_terms: D000595:Amino Acid Sequence; D001419:Bacteria / Q000201:enzymology; D004926:Escherichia coli / Q000201:enzymology*; D006714:Homoserine / Q000031:analogs & derivatives* / Q000378:metabolism; D007700:Kinetics; D008969:Molecular Sequence Data; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000737:chemistry* / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000201:enzymology; D017386:Sequence Homology, Amino Acid; D013050:Spectrometry, Fluorescence; D013379:Substrate Specificity
chemical_list: D006714:Homoserine; D017853:Phosphotransferases (Alcohol Group Acceptor); C021370:homoserine kinase