Molecular Modeling and Site Directed Mutagenesis of the O-Methyltransferase, SOMT-9 Reveal Amino Acids Important for Its Reaction and Regioselectivity

Bulletin of the Korean chemical society

Bong Gyu Kim¹  Yoongho Lim¹  So Hyun Park¹  Joong-Hoon Ahn¹  Sun Hee Lee¹  Youhoon Cheong¹ 

关键词: O-Methyltransferase;    Molecular modeling;    Quercetin;    Site-directed mutagenesis;   

DOI  :  

学科分类：化学（综合）

来源: Korean Chemical Society

【 摘 要 】

SOMT-9 is an O-methyltransferase that utilizes quercetin to produce 3`-methoxy quercetin. In order to determine which amino acids of SOMT-9 are important for this reaction and its regioselectivity, molecular docking experiments followed by site directed mutagenesis were performed. Molecular modeling and molecular docking experiments identified several amino acid residues involved in metal binding, AdoMet binding, and substrate binding. Site-directed mutagenesis showed that Asp188 is critical for metal binding and that Lys165 assists other metal binding residues in maintaining quercetin in the proper position during the reaction. In addition, Tyr207 was shown to play an important role in the determination of the regioselectivity and Met60 was shown to be involved in formation of the hydrophobic pocket necessary for substrate binding. The molecular modeling and docking experiments discussed in this study could be applicable to future research including prediction of substrate binding and regioselectivity of an enzyme.

【 授权许可】

Unknown