Partial purification and immunoreactivity of an 80 000-molecular-weight polypeptide associated with peroxisome proliferation in rat liver.
Partial purification and immunoreactivity of an 80 000-molecular-weight polypeptide associated with peroxisome proliferation in rat liver.
复制标题
与大鼠肝脏过氧化物酶体增殖相关的 80 000 分子量多肽的部分纯化和免疫反应性。
DOI：
10.1042/bj1880731
复制
发表时间：
1980
期刊：
影响因子：
--
通讯作者：
Reddy,JK
中科院分区：
文献类型：
--
作者：
Reddy,MK;Hollenberg,PF;Reddy,JK
降血脂药物和工业增塑剂，如邻苯二甲酸二（2-乙基己基）酯，这会导致肝过氧化物酶体的增殖，也会导致80000-mol.在大鼠和小鼠的肝脏中的多肽。这种多肽被命名为PPA-80（PPA，过氧化物酶体增殖相关的; 80代表80000mol.wt.）。用十二烷基硫酸钠/聚丙烯酰胺凝胶电泳一步纯化法，从用过氧化物酶体增殖剂Wy-14，643、萘非诺平、替布酸和氯贝特处理的大鼠肝脏中纯化出纯度超过90%的多肽PPA-80。从用Wy-14，643处理的大鼠的肝脏中分离的PPA-80多肽产生的抗体与从用Wy-14，643处理的大鼠的肝脏以及从用萘非诺平、替布酸和氯贝丁酯处理的大鼠的肝脏中纯化的多肽PPA-80发生交叉反应。抗-（多肽PPA-80）抗体不与过氧化氢酶（过氧化物酶体的标记酶）或NADPH-细胞色素P-450还原酶（其具有相同的近似摩尔重量）交叉反应，80000.与过氧化物酶体增殖剂预处理的动物肝脏的微粒体，大颗粒和核后组分形成的免疫荧光素带的强度显着大于从对照动物获得的相应组分的等量蛋白质相比，这表明这些试剂都增强了相同的80000-mol. wt.多肽。虽然多肽PPA-80的增加，在核后，大颗粒和微粒体组分的大鼠肝脏过氧化物酶体增殖剂预处理，这种肽的相对丰度的过氧化物酶体丰富的光线粒体组分和它的缺乏在高度纯化的线粒体组分表明本地化的这种多肽在过氧化物酶体和/或微粒体组分。需要另外的研究来明确地建立多肽PPA-80的亚细胞定位，并确定该多肽是否与通过Osumi和Hashimoto [（1979）Biochem. Biophys. Res. Commun.89，580 -584]。
Hypolipidaemic drugs and industrial plasticizers such as di-(2-ethylhexyl) phthalate, which cause proliferation of hepatic peroxisomes, also cause an increase in an 80000-mol.wt. polypeptide in the liver of rats and mice. This polypeptide has been designated as PPA-80 (PPA, for peroxisome-proliferation-associated; 80 for 80000mol.wt.). The polypeptide PPA-80 was purified to over 90% purity from livers of rats treated with the peroxisome proliferators Wy-14,643, nafenopin, tibric acid and clofibrate by a single-step preparative sodium dodecyl sulphate/polyacrylamide-gel-electrophoretic procedure. The antibodies raised against the PPA-80 polypeptide isolated from livers of rats treated with Wy-14,643 cross-reacted with polypeptide PPA-80 purified from the livers of rats treated with Wy-14,643, as well as from the livers of rats treated with nafenopin, tibric acid and clofibrate. The anti-(polypeptide PPA-80) antibodies did not cross-react with catalase, a marker enzyme for peroxisomes, or with NADPH–cytochromeP-450 reductase, which has the same approximate mol.wt., 80000. The intensity of immunoprecipitin bands formed with microsomal, large-particle and postnuclear fractions from livers of animals pretreated with peroxisome proliferators was significantly greater compared with equal amounts of protein from corresponding fractions obtained from control animals, suggesting that these agents all enhance the synthesis of the same 80000-mol.wt. polypeptide. Although the polypeptide PPA-80 was increased in the postnuclear, large-particle and microsomal fractions of livers of rats pretreated with peroxisome proliferators, the relative abundance of this peptide in the peroxisome-rich light-mitochondrial fraction and its lack in highly purified mitochondrial fractions suggest the localization of this polypeptide in peroxisomes and/or microsomal fraction. Additional studies are needed to establish unequivocally the subcellular localization of the polypeptide PPA-80 and to ascertain if this polypeptide is identical with the multi-functional protein displaying enoyl-CoA hydratase and β-hydroxyacyl-CoA dehydrogenase activities that was purified by Osumi & Hashimoto [(1979)Biochem. Biophys. Res. Commun.89,580–584].