波谱学杂志2017，Vol.34Issue(1)：1-7,7.DOI:10.11938/cjmr20170101
蛋白质INSM1中的锌指结构域ZF(4-5)的表达、纯化与表征
Expression, Purification and Characterization of the Zinc-Finger(4-5) Domain in Human Protein INSM1
摘要
Abstract
Human insulinoma-associated protein 1 (INSM1) is a transcriptional regulator recognizing sequence-specific DNA through its C-terminal zinc finger (ZF) domains. INSM1 contains five zinc finger domains, whose structures are still not known; therefore, the mechanisms through which it recognizes DNA also remain unclear. In this study, we designed the recombinant plasmid pET-32m-INSM1(424-497), which can express the truncated INSM1 fragment containing the zinc finger domains 4 and 5 [i.e., ZF(4-5)]. Expression and purification of ZF(4-5) were explored in order to achieve high protein yield for further structural and functional study. Nuclear magnetic resonance (NMR) and circular dichroism (CD) spectra revealed that chelation of Zn2+ to C2H2 in the ZF(4-5) was important for its structural stability, and also confirmed that the active-sites, Zn2+-chelated histidines, had the δ -tautomeric form.关键词
液体核磁共振(solutionNMR)/锌指蛋白/表达与纯化/结构与功能Key words
solution NMR/zinc finger domain/protein expression and purification/structure and function分类
数理科学引用本文复制引用
王华谱,朱勤俊,刘买利,杨运煌,岳霞丽..蛋白质INSM1中的锌指结构域ZF(4-5)的表达、纯化与表征[J].波谱学杂志,2017,34(1):1-7,7.基金项目
国家自然科学基金资助项目(21575155), 国家重点研发计划资助项目(2016YFA051201). （21575155）
