Yamamoto K, Völkl A, Fahimi H D
Department of Anatomy and Cell Biology II, University of Heidelberg, Germany.
J Histochem Cytochem. 1992 Dec;40(12):1909-18. doi: 10.1177/40.12.1360481.
We investigated the immunoreactivity of the peroxisomal lipid beta-oxidation enzymes acyl-CoA oxidase, trifunctional protein, and thiolase in guinea pig liver and compared it with that of homologous proteins in rat, using immunoblotting of highly purified peroxisomal fractions and monospecific antibodies to rat proteins. In addition, the immunocytochemical localization of beta-oxidation enzymes in guinea pig liver was compared with that of catalase. All antibodies showed crossreactivity between the two species, indicating that these peroxisomal proteins have been well conserved, although all exhibited some differences with respect to molecular size and, in the case of acyl-CoA oxidase, in frequency of the immunoreactive bands. In the latter case, a distinct second band in the 70 KD range was observed in guinea pig, in addition to the regular band due to subunit A present in rat liver. This novel band could be due either to trihydroxycoprostanoyl-CoA oxidase or to the non-inducible branched chain fatty acid oxidase described recently. All three beta-oxidation enzymes were immunolocalized by light and electron microscopy to the matrix of peroxisomes, in contrast to catalase, which is also found in the cytoplasm and the nucleus of hepatocytes in guinea pig liver.
我们利用高度纯化的过氧化物酶体组分的免疫印迹法以及针对大鼠蛋白的单特异性抗体，研究了豚鼠肝脏中过氧化物酶体脂质β-氧化酶酰基辅酶A氧化酶、三功能蛋白和硫解酶的免疫反应性，并将其与大鼠同源蛋白的免疫反应性进行了比较。此外，还比较了豚鼠肝脏中β-氧化酶与过氧化氢酶的免疫细胞化学定位。所有抗体在这两个物种之间均表现出交叉反应性，表明这些过氧化物酶体蛋白得到了很好的保守，尽管它们在分子大小方面都存在一些差异，对于酰基辅酶A氧化酶而言，免疫反应条带的频率也存在差异。在后一种情况下，除了大鼠肝脏中存在的由于亚基A产生的常规条带外，在豚鼠中还观察到了一条70 KD范围内明显的第二条带。这条新带可能是由于三羟胆甾烷酰辅酶A氧化酶或最近描述的非诱导性支链脂肪酸氧化酶所致。与过氧化氢酶不同，所有三种β-氧化酶通过光学显微镜和电子显微镜免疫定位到过氧化物酶体基质，在豚鼠肝脏中，过氧化氢酶也存在于肝细胞的细胞质和细胞核中。
