Structures and Protein Engineering of the α‐Keto Acid C‐Methyltransferases SgvM and MrsA for Rational Substrate Transfer

Structures and Protein Engineering of the α‐Keto Acid C‐Methyltransferases SgvM and MrsA for Rational Substrate Transfer

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 biosynthesi...

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Published in:Chembiochem : a European journal of chemical biology Vol. 25; no. 19; pp. e202400258 - n/a
Main Authors: Sommer‐Kamann, Christina, Breiltgens, Juliane, Zou, Ziruo, Gerhardt, Stefan, Saleem‐Batcha, Raspudin, Kemper, Florian, Einsle, Oliver, Andexer, Jennifer N., Müller, Michael
Format: Journal Article
Language:English
Published: Germany Wiley Subscription Services, Inc 01-10-2024
Subjects:
asymmetric methylation
biocatalysis
Biosynthesis
C-alkylation
Drug resistance
Lipophilic
Methionine
Methylation
Mutagenesis
Natural products
Protein engineering
Residues
Substrates
X-ray
C-alkylation
asymmetric methylation
Biocatalysis
X-ray
Mutagenesis
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Summary: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. The S‐Adenosyl‐l‐methionine‐dependent C‐methyltransferases SgvM and MrsA catalyze the stereoselective methylation of enolizable α‐keto acids in the biosynthesis of antibiotic natural products. By investigating key residues of the crystallized enzymes and rational engineering of MrsA, the substrate scope can be transferred to α‐keto acid substrates from different biosynthetic pathways, enabling enzymatic C‐methylation beyond native substrates.
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ISSN:1439-4227
1439-7633
1439-7633
DOI:10.1002/cbic.202400258