SETD2蛋白赖氨酸甲基转移酶的序列特异性分析及SETD2超级底物的发现。
Sequence specificity analysis of the SETD2 protein lysine methyltransferase and discovery of a SETD2 super-substrate.
作者信息
Schuhmacher Maren Kirstin, Beldar Serap, Khella Mina S, Bröhm Alexander, Ludwig Jan, Tempel Wolfram, Weirich Sara, Min Jinrong, Jeltsch Albert
机构信息
Institute of Biochemistry and Technical Biochemistry, University of Stuttgart, Allmandring 31, 70569, Stuttgart, Germany.
Structural Genomics Consortium, University of Toronto, 101 College Street, Toronto, ON, M5G 1L7, Canada.
出版信息
Commun Biol. 2020 Sep 16;3(1):511. doi: 10.1038/s42003-020-01223-6.
SETD2 catalyzes methylation at lysine 36 of histone H3 and it has many disease connections. We investigated the substrate sequence specificity of SETD2 and identified nine additional peptide and one protein (FBN1) substrates. Our data showed that SETD2 strongly prefers amino acids different from those in the H3K36 sequence at several positions of its specificity profile. Based on this, we designed an optimized super-substrate containing four amino acid exchanges and show by quantitative methylation assays with SETD2 that the super-substrate peptide is methylated about 290-fold more efficiently than the H3K36 peptide. Protein methylation studies confirmed very strong SETD2 methylation of the super-substrate in vitro and in cells. We solved the structure of SETD2 with bound super-substrate peptide containing a target lysine to methionine mutation, which revealed better interactions involving three of the substituted residues. Our data illustrate that substrate sequence design can strongly increase the activity of protein lysine methyltransferases.
SETD2催化组蛋白H3赖氨酸36位点的甲基化，且与多种疾病相关。我们研究了SETD2的底物序列特异性，确定了另外9种肽底物和1种蛋白质（FBN1）底物。我们的数据表明，在其特异性图谱的几个位置上，SETD2强烈偏好与H3K36序列中不同的氨基酸。基于此，我们设计了一种含有四个氨基酸替换的优化超级底物，并通过与SETD2的定量甲基化分析表明，超级底物肽的甲基化效率比H3K36肽高约290倍。蛋白质甲基化研究证实，在体外和细胞中，超级底物的SETD2甲基化作用非常强。我们解析了结合了含有靶赖氨酸到甲硫氨酸突变的超级底物肽的SETD2结构，该结构揭示了涉及三个取代残基的更好的相互作用。我们的数据表明，底物序列设计可以显著提高蛋白质赖氨酸甲基转移酶的活性。
Zotero 插件