Alam Nawsad, Zimmerman Lior, Wolfson Noah A, Joseph Caleb G, Fierke Carol A, Schueler-Furman Ora
Department of Microbiology and Molecular Genetics, Institute for Medical Research Israel-Canada, Faculty of Medicine, Hebrew University of Jerusalem, Jerusalem 91120, Israel.
Department of Biological Chemistry, University of Michigan, Ann Arbor, MI 48109-1055, USA.
Structure. 2016 Mar 1;24(3):458-68. doi: 10.1016/j.str.2016.02.002.
HDAC8 is a member of the family of histone deacetylases (HDACs) that catalyze the deacetylation of acetyl lysine residues within histone and non-histone proteins. The recent identification of novel non-histone HDAC8 substrates such as SMC3, ERRα, and ARID1A indicates a complex functionality of this enzyme in cellular homeostasis. To discover additional HDAC8 substrates, we developed a comprehensive, structure-based approach based on Rosetta FlexPepBind, a protocol that evaluates peptide-binding ability to a receptor from structural models of this interaction. Here we adapt this protocol to identify HDAC8 substrates using peptide sequences extracted from proteins with known acetylated sites. The many new in vitro HDAC8 peptide substrates identified in this study suggest that numerous cellular proteins are HDAC8 substrates, thus expanding our view of the acetylome and its regulation by HDAC8.
HDAC8是组蛋白去乙酰化酶（HDACs）家族的成员，可催化组蛋白和非组蛋白中乙酰赖氨酸残基的去乙酰化反应。最近发现的新型非组蛋白HDAC8底物，如SMC3、ERRα和ARID1A，表明该酶在细胞内稳态中具有复杂的功能。为了发现更多HDAC8底物，我们基于Rosetta FlexPepBind开发了一种全面的、基于结构的方法，该方法可从这种相互作用的结构模型评估肽与受体的结合能力。在这里，我们采用此方法，利用从具有已知乙酰化位点的蛋白质中提取的肽序列来鉴定HDAC8底物。本研究中鉴定出的许多新的体外HDAC8肽底物表明，众多细胞蛋白都是HDAC8底物，从而拓展了我们对乙酰化蛋白质组及其受HDAC8调控的认识。
