Cheng Dong, Chu Ching-Hsuen, Chen Luping, Feder John N, Mintier Gabe A, Wu Yuli, Cook Joseph W, Harpel Mark R, Locke Gregory A, An Yongmi, Tamura James K
Department of Obesity and Metabolic Research, Pharmaceutical Research Institute, Bristol-Myers Squibb Company, PO Box 5400, Princeton, NJ 08543-5400, USA.
Protein Expr Purif. 2007 Jan;51(1):11-21. doi: 10.1016/j.pep.2006.06.005. Epub 2006 Jun 10.
Acetyl coenzyme A (acetyl-CoA) carboxylase isozyme 1 (ACC1) and acetyl-CoA carboxylase isozyme 2 (ACC2) are critical for de novo fatty acid synthesis and for the regulation of beta-oxidation. Emerging evidence indicates that one or both isozymes might be therapeutic targets for the treatment of obesity, type 2 diabetes, and dyslipidemia. One of the major obstacles in the field is the lack of readily-available source of recombinant human ACC enzymes to support systematic drug discovery efforts. Here, we describe an efficient and optimal protocol for expressing and isolating recombinant mammalian ACCs with high yield and purity. The resultant human ACC2, human ACC1, and rat ACC2 possess high specific activities, are properly biotinylated, and exhibit kinetic parameters very similar to the native ACC enzymes. We believe that the current study paves a road to a systematic approach for drug design revolving around the ACC inhibition mechanism.
乙酰辅酶A（acetyl-CoA）羧化酶同工酶1（ACC1）和乙酰辅酶A羧化酶同工酶2（ACC2）对于从头脂肪酸合成以及β-氧化的调节至关重要。新出现的证据表明，一种或两种同工酶可能是治疗肥胖症、2型糖尿病和血脂异常的治疗靶点。该领域的主要障碍之一是缺乏易于获得的重组人ACC酶来源，以支持系统性药物研发工作。在此，我们描述了一种高效且优化的方案，用于高产率和高纯度地表达和分离重组哺乳动物ACC。所得的人ACC2、人ACC1和大鼠ACC2具有高比活性，能被正确生物素化，并且表现出与天然ACC酶非常相似的动力学参数。我们相信，当前的研究为围绕ACC抑制机制进行药物设计的系统性方法铺平了道路。
