酵母Efm4或人类eEF1A-KMT2对延长因子1A的甲基化涉及β-hairpin识别动机,并与酸化交叉
Joshua J Hamey1, Amy Nguyen1, Mahdi Haddad1
1School of Biotechnology and Biomolecular Sciences, University of New South Wales, New South Wales, Australia.
The Journal of biological chemistry
|January 10, 2024
概括
这项研究揭示了Efm4甲基转移酶如何在K316使用独特的β-hairpin动机选择性地修改K316的翻译延长因子1A (eEF1A). 这种机制解释了蛋白质甲基化中的酶选择性.
科学领域:
- 生物化学 生物化学
- 分子生物学分子生物学
- 蛋白质组学是指蛋白质组学.
背景情况:
- 翻译延长因子1A (eEF1A) 对于蛋白质合成至关重要,并被多种酶广泛甲基化.
- 对于eEF1A甲基转移酶对特定残留物的高选择性仍然不明.
研究的目的:
- 阐明Saccharomyces cerevisiae延长因子甲基转移酶4 (Efm4) 在lysine 316 (K316) 中选择性地甲基化eEF1A的机制.
主要方法:
- 使用AlphaFold-Multimer建模,交叉连接质谱 (XL-MS) 和酶突变发生.
- 研究了β-hairpin图案在Efm4及其人类同类METTL10.0中的作用.
- 评估了eEF1Bα结合和eEF1A酸化对甲基化活性的影响.
主要成果:
- 在Efm4中有一个独特的β-hairpin图案,对eEF1A K316甲基化至关重要.
- 在Efm4中突变氨酸212 (F212) 显著降低了其在体外和酵母中的活性.
- 人类METTL10中相当于F220的残留物对eEF1A甲基化也很重要.
- eEF1Bα竞争性地抑制了Efm4的活动,eEF1A S314酸化与K316甲基化负面交叉.
结论:
- 蛋白甲基转移酶通过特定的结构动机 (如β-hairpin) 实现高选择性.
- 确定了控制Efm4-eEF1A甲基化特异性的关键残留物和调控因素 (eEF1Bα,酸化).
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