Elucidation of activation mechanism of human acetyl-CoA carboxylase

阐明人乙酰辅酶A羧化酶的激活机制

• 批准号: 22590113
• 负责人: KUNISHIMA Naoki
• 金额: $ 2.08万
• 依托单位: The Institute of Physical and Chemical Research
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2010
• 资助国家: 日本
• 起止时间: 2010 至 2012
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-22590113/
• 关键词: 生活習慣病; 脂肪酸生合成; 結晶構造解析; X線回折; 酵素; タンパク質; 立体構造; 薬物設計

Toward the elucidation of the activation mechanism of acetyl-CoA carboxylase (ACC) that catalyzes the rate-limiting step of the fatty-acid biosynthesis, we tried the protein crystallography of the central domain of human ACC (hBCCP1, hBCCP2) and its activation enzyme biotin protein ligase (hBPL). The genes were synthesized for a large-scale expression using E.coli. These proteins were generally expressed as inclusion bodies that could not be solubilized by methods including the denaturation/refolding and the substitution of cysteine with alanine.

为了阐明催化脂肪酸生物合成限速步骤的乙酰辅酶A羧基酶(ACC)的激活机制，我们尝试了人ACC中央结构域的蛋白质结晶学(hBCCP1，hBCCP2)及其激活酶生物素蛋白连接酶(HBPL)。这些基因是用大肠杆菌合成的，可以大规模表达。这些蛋白通常以包涵体的形式表达，不能通过变性/复性和半胱氨酸替代丙氨酸等方法溶解。