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

FEBS Letters

Selvapandiyan, Angamuthu¹  Arora, Naresh¹  Ahmad, Suhail¹  Bhatnagar, Raj K.¹  Fattah, Farkad A.¹  Majumder, Kumud¹ 

[1] International Centre for Genetic Engineering and Biotechnology, N.I.L. Campus, Aruna Asaf Ali Marg, New Delhi 110 067, India

关键词: 5-Enolpyruvyl shikimate 3-phosphate synthase;    Site-directed mutagenesis;    Active site analysis;    Glyphosate;   

DOI  :  10.1016/0014-5793(95)01124-W

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

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.

【 授权许可】

Unknown