Expression, purification, and characterization of human and rat acetyl coenzyme A carboxylase (ACC) isozymes
Expression, purification, and characterization of human and rat acetyl coenzyme A carboxylase (ACC) isozymes
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DOI：
10.1016/j.pep.2006.06.005
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发表时间：
2007-01-01
影响因子：
1.6
通讯作者：
Tamura, James K.
中科院分区：
文献类型：
--
作者：
Cheng, Dong;Chu, Ching-Hsuen;Tamura, James K.
乙酰辅酶 A (乙酰辅酶 A) 羧化酶同工酶 I (ACC1) 和乙酰辅酶 A 羧化酶同工酶 2 (ACC2) 对于脂肪酸从头合成和 β-氧化调节至关重要。新的证据表明，一种或两种同工酶可能是治疗肥胖、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现成的重组人 ACC 酶来源来支持系统的药物发现工作。这里。我们描述了一种高效且最佳的方案，用于以高产量和纯度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，经过适当的生物素化，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 抑制机制的药物设计系统方法铺平了道路。 (c) 2006 Elsevier Inc. 保留所有权利。
Acetyl coenzyme A (acetyl-CoA) carboxylase isozyme I (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. (c) 2006 Elsevier Inc. All rights reserved.