Friedrich Miescher Institute for Biomedical Research, Basel, Switzerland.
Faculty of Sciences, University of Basel, Basel, Switzerland.
Methods Mol Biol. 2023;2589:467-480. doi: 10.1007/978-1-0716-2788-4_30.
Histone deacetylase 6 (HDAC6) is an atypical lysine deacetylase with tandem catalytic domains and an ubiquitin-binding zinc finger domain. HDAC6 is involved in various biological processes, such as cell motility or stress responses, and has been implicated in pathologies ranging from cancer to neurodegeneration. Due to this broad range of functions, there has been considerable interest in developing HDAC6-specific small molecule inhibitors, several of which are already available. The crystal structure of the tandem catalytic domains of zebrafish HDAC6 has revealed an arrangement with twofold symmetry and extensive surface interaction between the catalytic domains. Further dissection of the biochemical properties of HDAC6 and the development of novel inhibitors will benefit from being able to routinely express high-quality protein. We present here our optimized protocol for expression and crystallization of the zebrafish tandem catalytic domains.
组蛋白去乙酰化酶 6（HDAC6）是一种具有串联催化结构域和泛素结合锌指结构域的非典型赖氨酸去乙酰化酶。HDAC6 参与多种生物学过程，如细胞运动或应激反应，并与从癌症到神经退行性变等各种病理学有关。由于其广泛的功能，人们对开发 HDAC6 特异性小分子抑制剂产生了浓厚的兴趣，其中一些已经问世。斑马鱼 HDAC6 串联催化结构域的晶体结构揭示了具有两倍对称性的排列方式和催化结构域之间广泛的表面相互作用。对 HDAC6 的生化特性进行进一步剖析并开发新型抑制剂，将得益于能够常规表达高质量的蛋白质。我们在此介绍优化的斑马鱼串联催化结构域表达和结晶方案。
