Global Profiling of 2-hydroxyisobutyrylome in Common Wheat.
Global Profiling of 2-hydroxyisobutyrylome in Common Wheat.
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普通小麦 2-羟基异丁酰组的整体分析
DOI：
10.1016/j.gpb.2020.06.008
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发表时间：
2022-08
影响因子：
9.5
通讯作者：
Chen, Feng
中科院分区：
文献类型：
--
作者：
Zhang, Ning;Zhang, Lingran;Li, Linjie;Geng, Junyou;Zhao, Lei;Ren, Yan;Dong, Zhongdong;Chen, Feng
关键词：
作为一种新的翻译后修饰(PTM)，赖氨酸2-羟基异丁酰化(KHIB)被认为在真核细胞中调节基因的转录活性；然而，KHIB修饰的蛋白在植物中的功能尚不清楚。在此，我们报道了Khib是小麦及其祖先中一种进化保守的PTM。普通小麦(Triticum aestivum L.)的1074个蛋白质上共有3348个Khib位点。2-羟基异丁烯的亲和纯化和质谱分析。生物信息学数据表明，Khib修饰的蛋白质参与了广泛的生物和代谢途径。免疫沉淀证实Khib修饰的蛋白质是内源性存在的。Khib与其他主要PTM的比较表明，Khib修饰的蛋白质同时被多个PTMS修饰。通过突变实验和免疫共沉淀实验，我们证明了磷酸甘油酸激酶(PGK)K206上的Khib是其酶活性的关键调控修饰，K206上的突变影响了PGK与其底物的相互作用。此外，低分子蛋白质的Khib修饰是对脱乙酰酶抑制剂烟酰胺和曲古菌素的反应。这项研究为我们目前对小麦植株中Khib的了解提供了证据，包括Khib与其代谢调控之间的合作。
As a novel post-translational modification (PTM), lysine 2-hydroxyisobutyrylation (Khib) is considered to regulate gene transcriptional activities in eukaryotic cells; however, the functions of Khib-modified proteins in plants remain unknown. Here, we report that Khib is an evolutionarily-conserved PTM in wheat and its progenitors. A total of 3348 Khib sites on 1074 proteins are identified in common wheat (Triticum aestivum L.) by using affinity purification and mass spectroscopy of 2-hydroxyisobutyrylome. Bioinformatic data indicate that Khib-modified proteins participate in a wide variety of biological and metabolic pathways. Immunoprecipitation confirms that Khib-modified proteins are present endogenously. A comparison of Khib and other main PTMs shows that Khib-modified proteins are simultaneously modified by multiple PTMs. Using mutagenesis experiments and co-immunoprecipitation assays, we demonstrate that Khib on K206 of phosphoglycerate kinase (PGK) is a key regulatory modification for its enzymatic activity, and mutation on K206 affects the interactions of PGK with its substrates. Furthermore, Khib modification of low-molecular-weight proteins is a response to the deacetylase inhibitors nicotinamide and trichostatin. This study provides evidence to promote our current understanding of Khib in wheat plants, including the cooperation between Khib and its metabolic regulation.
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