title: Mutational analysis of the CitA citrate transporter from Salmonella typhimurium: altered substrate specificity.
abstract: The CitA citrate transporter in Salmonella typhimurium is encoded by the citA gene and consists of 434 amino acid residues that probably include 12 membrane-spanning segments [Shimamoto. T., et al. (1991) J. Biochem. 110, 22-28]. CitA mutants with altered substrate specificities were isolated by in vitro mutagenesis using nitrous acid. The mutants could grow on isocitrate as a sole carbon source which normally cannot be transported well by the CitA transporter of S. typhimurium. The mutation sites in the citA gene of the nine mutants were determined to involve single residues at seven sites (one mutation per mutant). The original amino acid residues at these sites (Arg-19, Ala-38, Glu-51, Gly-132, Ala-169, Pro-262 and Leu-271) were identified to be responsible for the altered substrate specificity. All these amino acid residues were conserved in four other homologous citrate transporters from Escherichia coli, Citrobacter amalonaticus and Klebsiella pneumoniae and are suggested to be involved in substrate recognition by the CitA transporter.
authors.affiliation: Gene Research Center, Faculty of Pharmaceutical Sciences, Okayama University, Japan.
authors.forename: T
authors.initials: T
authors.lastname: Shimamoto
authors.forename: K
authors.initials: K
authors.lastname: Negishi
authors.forename: M
authors.initials: M
authors.lastname: Tsuda
authors.forename: T
authors.initials: T
authors.lastname: Tsuchiya
mesh_terms: D000595:Amino Acid Sequence; D002352:Carrier Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D011487:Protein Conformation; D012486:Salmonella typhimurium / Q000235:genetics*; D016415:Sequence Alignment; D013379:Substrate Specificity
chemical_list: D002352:Carrier Proteins; C020723:citrate-binding transport protein