Henningfield M F, Swick R W
Biochem Cell Biol. 1987 Mar;65(3):245-51. doi: 10.1139/o87-032.
A polyclonal antisera against rat brown adipose tissue mitochondrial uncoupling protein was used to examine mitochondrial samples from liver and white and brown adipose tissue from several mammalian species. A sodium dodecyl sulfate--polyacrylamide gel electrophoretic separation of proteins combined with an immunochemical method allowed for visualization of antigen--antibody complexes on nitrocellulose blots. Hamster, cavy, monkey, and mouse brown adipose tissue mitochondrial samples cross-reacted with the antisera. Mitochondria prepared from white fat obtained from young swine and sheep contained two closely migrating, antigenically active proteins. Hepatic mitochondria samples did not contain antigenically active protein. Reflectance densitometry was used for quantitation of the uncoupling protein in various mitochondrial samples. In rats fed diets low in protein, there appears to be a dissociation between the concentration of uncoupling protein and the number of nucleotide binding sites as given by the [3H]GDP binding assay. These results are indicative of a physiological activation of the uncoupling protein.
使用针对大鼠棕色脂肪组织线粒体解偶联蛋白的多克隆抗血清，检测了来自几种哺乳动物物种的肝脏、白色和棕色脂肪组织的线粒体样本。蛋白质的十二烷基硫酸钠-聚丙烯酰胺凝胶电泳分离与免疫化学方法相结合，使得能够在硝酸纤维素印迹上可视化抗原-抗体复合物。仓鼠、豚鼠、猴子和小鼠的棕色脂肪组织线粒体样本与抗血清发生交叉反应。从幼猪和绵羊的白色脂肪中制备的线粒体含有两种迁移紧密、具有抗原活性的蛋白质。肝脏线粒体样本不含具有抗原活性的蛋白质。反射密度测定法用于定量各种线粒体样本中的解偶联蛋白。在喂食低蛋白饮食的大鼠中，解偶联蛋白的浓度与通过[3H]GDP结合试验测定的核苷酸结合位点数量之间似乎存在解离。这些结果表明解偶联蛋白发生了生理激活。
