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对于合理基质转移的α-基托酸C-甲基转移酶SgvM和MrsA的结构和蛋白质工程

Christina Sommer-Kamann¹, Juliane Breiltgens¹, Ziruo Zou¹

¹Institute of Pharmaceutical Sciences, University of Freiburg, Albertstrasse 25, 79104, Freiburg, Germany.

Chembiochem : a European journal of chemical biology

|June 18, 2024

概括

研究人员设计了一种甲基转移酶 (MT) 酶,MrsA,以接受更广泛的天然产品基质. 这项工作展示了合理的设计,用于在不同的α-基托酸MT之间转移基质范围.

科学领域:

生物化学生物化学
酶学是一种酶学.
自然产品生物合成自然产品生物合成

背景情况:

依赖S-adenosyl-l-methionine的甲基转移酶 (MTs) 对于通过C-甲基化修改自然产品至关重要.
具体的MTs,SgvM和MrsA,甲基酸α-keto酸分别在维里多格里塞因和3-甲基亚尔金因的生物合成中.
与SgvM不同,MrsA表现出严格的基质选择性.

研究的目的:

在SgvM和MrsA中阐明基质认可的结构基础.
为扩大基板范围设计MrsA,接受SgvM基板.
通过合理设计来证明α-酸MT之间的基质范围的转移.

主要方法:

用X射线结晶学测定SgvM和MrsA的apo和基质/辅因子结合结构.
位点定向的突变发生改变MrsA的关键活性位点残留物.
生物化学试验,以评估野生类型和人工MrsA的基质接受度.

主要成果:

晶体结构揭示了SgvM和MrsA活性位点中控制基质特异性的关键残留物.
工程设计的MrsA变体证明了对SgvMα-keto酸基质的接受,其中含有未充电和脂性β-残留物.
该研究成功地将基质识别能力转移到不同的MT酶之间.

结论:

对SgvM和MrsA活性部位的结构洞察力有助于酶工程.
合理修改MrsA扩大了其基质范围,使其能够合成各种甲基化天然产品.
这项工作为新型生物催化应用的工程MT提供了一个框架.

关键词:

在C-基化过程中,这是X射线.不对称的甲基化.生物催化剂的生物催化剂突变发生的突变发生.

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