title: Point mutation of a conserved arginine (104) to lysine introduces hypersensitivity to inhibition by glyphosate in the 5-enolpyruvylshikimate-3-phosphate synthase of Bacillus subtilis.
abstract: The role of a conserved arginine (R104) in the putative phosphoenol pyruvate binding region of 5-enolpyruvyl shikimate-3-phosphate synthase of Bacillus subtilis has been investigated. Employing site directed mutagenesis arginine was substituted by lysine or glutamine. Native and mutant proteins were expressed and purified to near homogeneity. Estimation of Michaelis and inhibitor constants of the native and mutant proteins exhibited altered substrate-inhibitor binding mode and constants. Mutation R104K hypersensitized the enzyme reaction to inhibition by glyphosate. The role of R104 in discriminating between glyphosate and phosphoenol pyruvate is discussed.
authors.affiliation: International Centre for Genetic Engineering and Biotechnology, Aruna Asaf Ali Marg, New Delhi, India.
authors.forename: A
authors.initials: A
authors.lastname: Selvapandiyan
authors.forename: K
authors.initials: K
authors.lastname: Majumder
authors.forename: F A
authors.initials: FA
authors.lastname: Fattah
authors.forename: S
authors.initials: S
authors.lastname: Ahmad
authors.forename: N
authors.initials: N
authors.lastname: Arora
authors.forename: R K
authors.initials: RK
authors.lastname: Bhatnagar
mesh_terms: D051229:3-Phosphoshikimate 1-Carboxyvinyltransferase; D019883:Alkyl and Aryl Transferases; D000595:Amino Acid Sequence; D001120:Arginine / Q000737:chemistry*; D001412:Bacillus subtilis / Q000201:enzymology* / Q000235:genetics; D001665:Binding Sites; D017124:Conserved Sequence; D004791:Enzyme Inhibitors / Q000737:chemistry / Q000494:pharmacology*; D005973:Glutamine / Q000737:chemistry; D005998:Glycine / Q000031:analogs & derivatives* / Q000737:chemistry / Q000494:pharmacology; D008239:Lysine / Q000737:chemistry; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D010728:Phosphoenolpyruvate / Q000378:metabolism; D017354:Point Mutation; D012765:Shikimic Acid / Q000031:analogs & derivatives / Q000378:metabolism; D013379:Substrate Specificity; D014166:Transferases / Q000037:antagonists & inhibitors* / Q000235:genetics; D000097797:Glyphosate
chemical_list: D051229:3-Phosphoshikimate 1-Carboxyvinyltransferase; D019883:Alkyl and Aryl Transferases; D001120:Arginine; D004791:Enzyme Inhibitors; D005973:Glutamine; D005998:Glycine; D008239:Lysine; D010728:Phosphoenolpyruvate; D012765:Shikimic Acid; D014166:Transferases; D000097797:Glyphosate; C042786:shikimic acid-3-phosphate