Cleeter M W, Ragan C I
Biochem J. 1985 Sep 15;230(3):739-46. doi: 10.1042/bj2300739.
The polypeptide composition of isolated mitochondrial NADH:ubiquinone reductase (NADH dehydrogenase) is very similar to that of material immunoprecipitated from detergent-solubilized bovine heart submitochondrial particles by antisera to the holoenzyme. The specificity of the antisera for dehydrogenase polypeptides was determined by immunoblotting, which showed that antisera reacting with only a few proteins were able to immunoprecipitate all others in parallel. The polypeptide compositions of rat, rabbit and human NADH dehydrogenase were determined by immunoprecipitation of the enzyme from solubilized submitochondrial particles and proved to be very similar to that of the bovine heart enzyme, particularly in the high-Mr region. Further homologies in these and other species were explored by immunoblotting with antisera to the holoenzyme and monospecific antisera raised against iron-sulphur-protein subunits of the enzyme.
分离得到的线粒体NADH：泛醌还原酶（NADH脱氢酶）的多肽组成，与用全酶抗血清从去污剂增溶的牛心亚线粒体颗粒中免疫沉淀得到的物质非常相似。通过免疫印迹法确定了抗血清对脱氢酶多肽的特异性，结果表明，只与少数几种蛋白质发生反应的抗血清能够同时免疫沉淀所有其他蛋白质。通过从增溶的亚线粒体颗粒中免疫沉淀该酶，测定了大鼠、兔子和人类NADH脱氢酶的多肽组成，结果证明它们与牛心酶的多肽组成非常相似，尤其是在高分子量区域。用全酶抗血清和针对该酶铁硫蛋白亚基产生的单特异性抗血清进行免疫印迹，进一步探索了这些物种和其他物种之间的同源性。
