中文摘要：

目的：寻找提取纯化具有12次跨膜结构的葡萄糖转运蛋白4（GLUT4）的新方法,并通过晶体结构的解析来研究二型糖尿病的发生机制。方法：首次选用猪肉皮下脂肪作为GLUT4蛋白提取的原材料,采用组织匀浆,差速离心方法初步提取分离了富含GLUT4的组分,进一步通过硫酸铵沉淀法初步纯化到大量的GLUT4蛋白。结果：通过单抗1F8对每一个组分进行Westem Blot检测,证明了分子量55kD处的蛋白即是GLUT4。150g脂肪组织通过组织匀浆和差速离心后,于35%-55%的硫酸铵沉淀浓度范围内得到40mg纯度达到50%的GLUT4。纯化后的GLUT4可以在0.5%DM下稳定存在三天。结论：发展了快速有效提取GLUT4的新方法,为进一步蛋白结晶打下基础。

英文摘要：

Objective： To find a new way of extraction and purification of Glucose Transporter 4 （GLUT4）, which have 12 trans-membrane segments, and study the mechanism of type Ⅱ diabetes by resolution of the 3D structure of GLUT4. Methods： We for the first time used fat tissues as the native material to harvest GLUT4. High yield of GLUT4 was extracted and purified from fat tissues by methods of homogenate, differential centrifugation and ammonium sulfate precipitation. Results： The purified GLUT4 with molecular weight of 55 kDa was verified by Western Blot with monoclonal antibody 1F8, after homogenate and differential centrifugation of 150g fat tissue, 40mg GLUT4 was obtained at 500/0 purity in the 35%-55% of ammonium sulfate solution. The purified GLUT4 protein can be stabilized in 0.5% DM for at least 3 days. Conclusion： The paper developed a method of effective purification of GLUT4 from fat dssue for further protein crystaUizadon.