1. Structures and Protein Engineering of the α-Keto Acid C-Methyltransferases SgvM and MrsA for Rational Substrate Transfer.
- Author
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Sommer-Kamann C, Breiltgens J, Zou Z, Gerhardt S, Saleem-Batcha R, Kemper F, Einsle O, Andexer JN, and Müller M
- Subjects
- Substrate Specificity, Pseudomonas syringae enzymology, Models, Molecular, Crystallography, X-Ray, Mutagenesis, Site-Directed, Bacterial Proteins metabolism, Bacterial Proteins genetics, Bacterial Proteins chemistry, Catalytic Domain, Methicillin-Resistant Staphylococcus aureus enzymology, Methyltransferases metabolism, Methyltransferases chemistry, Methyltransferases genetics, Protein Engineering, Streptomyces enzymology, Streptomyces genetics, Keto Acids metabolism, Keto Acids chemistry
- Abstract
S-adenosyl-l-methionine-dependent methyltransferases (MTs) are involved in the C-methylation of a variety of natural products. The MTs SgvM from Streptomyces griseoviridis and MrsA from Pseudomonas syringae pv. syringae catalyze the methylation of the β-carbon atom of α-keto acids in the biosynthesis of the antibiotic natural products viridogrisein and 3-methylarginine, respectively. MrsA shows high substrate selectivity for 5-guanidino-2-oxovalerate, while other α-keto acids, such as the SgvM substrates 4-methyl-2-oxovalerate, 2-oxovalerate, and phenylpyruvate, are not accepted. Here we report the crystal structures of SgvM and MrsA in the apo form and bound with substrate or S-adenosyl-l-methionine. By investigating key residues for substrate recognition in the active sites of both enzymes and engineering MrsA by site-directed mutagenesis, the substrate range of MrsA was extended to accept α-keto acid substrates of SgvM with uncharged and lipophilic β-residues. Our results showcase the transfer of the substrate scope of α-keto acid MTs from different biosynthetic pathways by rational design., (© 2024 The Authors. ChemBioChem published by Wiley-VCH GmbH.)
- Published
- 2024
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