Functional and Structural Analyses of trans C-Methyltransferase in Fungal Polyketide Biosynthesis
Functional and Structural Analyses of trans C-Methyltransferase in Fungal Polyketide Biosynthesis
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DOI：
10.1021/acs.biochem.9b00702
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发表时间：
2019-09-24
期刊：
影响因子：
2.9
通讯作者：
Watanabe, Kenji
中科院分区：
文献类型：
--
作者：
Kishimoto, Shinji;Tsunematsu, Yuta;Watanabe, Kenji
某些真菌聚酮化合物-肽合成酶的生物合成涉及C-甲基转移酶活性，其将一个或多个S-腺苷-L-甲硫氨酸衍生的甲基添加到碳框架上。先前报道的 PsoF-MT 是来自 pseurotin 生物合成途径的独立 C-甲基转移酶 (MT)，作为三功能双结构域酶 PsoF 中的一个结构域存在，使用具有底物类似物的突变体对晶体学和动力学进行了表征，以了解交易 C-MT 如何工作，并将其与已知的聚酮合酶相关 C-MT 进行比较。这项研究确定了参与催化和底物识别的关键活性位点残基，这使我们提出了 PsoF-MT 中 C-甲基化和底物特异性决定因素的机制。
Biosynthesis of certain fungal polyketide-peptide synthetases involves C-methyltransferase activity that adds one or more S-adenosyl-L-methionine-derived methyl groups to the carbon framework. The previously reported PsoF-MT, the stand-alone C-methyltransferase (MT) from the pseurotin biosynthetic pathway that exists as a domain within a trifunctional didomain enzyme PsoF, was characterized crystallographically and kinetically using mutants with substrate analogs to understand how a transacting C-MT works and compare it to known polyketide synthase-associated C-MTs. This study identified key active-site residues involved in catalysis and substrate recognition, which led us to propose the mechanism of C-methylation and substrate specificity determinants in PsoF-MT.