2012-03-012024-05-17https://scholars.lib.ntu.edu.tw/handle/123456789/681294摘要：人類細胞之組蛋白去乙醯&#37238;6 (Histone deacetylase 6, HDAC6) 扮演調節微管蛋白的乙醯化及去乙醯化，為一唯一存在於細胞質中。HDAC6 經確認在細胞遭遇不正常蛋白聚集時，造成神經退化的現象。有數種HDAC6的抑制劑已推測可作為神經退化疾病及癌症之用。因此，我們將探究HDAC6與這些抑制劑之交互作用，同時進一步設計及修改抑制劑之官能基，以進一步改善抑劑劑之效果。 為了解HDAC6及抑制劑間之結合，我們將使用生物物理研究方法去探究其分子功能。首先利用x光晶體繞射決定HDAC6之蛋白質晶體結構，同時與合成之quinazolinedione 衍生物進行複合體結構分析。第二、利用石英晶體微天秤及等溫淍定熱力儀決定這些衍生物與HDAC6之結合常數。第三、以結構生物資訊分析及靜電力計算提供分子表面電性資訊。最後，將能決定這些衍生物與HDAC6之結合官能基，並進一步設計更有效的衍生複合物，以作為未來藥物篩選之用。 為了取得純化的HDAC6重組蛋白，我們將在第一年構築hdac6基因進入表現載體，如PET21B 或pGEX4T1，進一步測試其表現及純化策略。在第二年，以純化重組HDAC6蛋白，利用共結晶的方法與其抑制劑(J22352, J22389 and J22390)一起培養，同時決定HDAC6與抑制劑之結合常數。第三年根據HDAC6與抑制劑之蛋白結構，設計具更佳結合效率抑制劑。據此，將可提供有力的分子證據發展HDAC6之小分子抑制劑藥物。 細胞質中組蛋白去乙醯&#37238;調節是一種重要的細胞事件，特別是在神經發育中。決定HDAC6-抑制劑複合體結構，將可提供更專一的小分子藥物抑制HDAC6之活性。它將可作為具有潛力發展神經藥物的方向之一。 <br> Abstract: Histone deacetylase 6 (HDAC6) in human cells is an enzyme to regulate tubulin acetylation and deacetylation, and is localized exclusively in the cytoplasm. HDAC6 has been identified to involve in neurodegeneration while cell encountered abnormal protein aggregation. Several inhibitors of HDAC6 are proposed in treatment of neurodegenerative disorders and cancer. Here, we would like to investigate the interaction between HDAC6 and inhibitors which will provide detailed binding properties to further design and modify the functional group of inhibitors for fitting and improving the inhibitor’s abilities. For revealing the interaction between HDAC6 and inhibitors, we aim to use several biophysics methods to explore the molecular mechanism. First, x-ray crystallography will use to determine the crystal structure of HDAC6 in complex with synthetic quinazolinedione derivative compounds from Professor Ji-Wang Chern. Second, quartz crystal microbalance and isothermal titration calorimetry can provide the binding constants between HDAC6 and the compounds. Third, structural bioinformatics and electrostatic calculation will provide the information of molecular surface. Finally, we could determine the functional groups of the compound to interact with HDAC6 and design higher or more efficacies of derivative compounds for collaborators in the research group. In order to obtain the purified HDAC6 protein, we aim to construct hdac6 gene into expression vectors, e.g. PET21b or pGEX4T1, and test its expression and purification strategies in the first year. The purified recombinant HDAC6 will use to co-crystallize with the inhibitors (J22352, J22389 and J22390 from Professor Ji-Wang Chern). The binding constants of HDAC6 with the inhibitors will also be determined in the second year. According to the structure of HDAC6 and the inhibitors complexes, the binding constants between HDAC6 and its inhibitors, the structure-based compound design will provide solid molecular evidence to develop small molecule inhibitors for HDAC6. The regulation of histone deacetylase in cytoplasm is an important cellular event, especially in neuron development. To resolve HDAC6-inhibitors complexes will provide more specific small molecules to inhibit the activity of HDAC6. It might be a potential direction to develop neurite drugs.HDAC6-抑制劑複合體晶體結構結合常數QCMHDAC6-inhibitors complexcrystal structurebinding constantsQCM