Häyrinen H M, Svensson L T, Hultenby K, Sormunen R T, Wilcke M, Hiltunen J K, Alexson S E
Biocenter, Oulu University, Finland.
Eur J Cell Biol. 1997 Jan;72(1):70-8.
Recently it has been shown that peroxisomes interact with microtubules which affect the structure and intracellular distribution of the organelle (Schrader, M., J. K. Burkhardt, E. Baumgart, G. Lüers, H. Spring, A. Völkl, H. D. Fahimi: Interaction of microtubules with peroxisomes. Tubular and spherical peroxisomes in HepG2 cells and their alterations induced by microtubule-active drugs. Eur. J. Cell Biol. 69, 24-35 (1996)). In the present work, we have applied immunological techniques to study the organization of peroxisomes within the rat liver cell. Antibodies to a pentadecapeptide corresponding to amino acid residues 403-417 of the 70 kDa integral peroxisomal membrane protein (Kamijo, K., S. Taketani, S. Yokota, T. Osumi, T. Hashimoto: The 70-kDa peroxisomal membrane protein is a member of the Mdr (P-glycoprotein)-related ATP-binding protein super family. J. Biol. Chem. 265, 4534-4540 (1990)) were raised in rabbits and affinity purified. This antibody was found to be highly specific for peroxisomes as determined by ELISA and Western blot analysis. Immunoelectron microscopy of tissue sections from rat liver revealed that peroxisomal membranes were labeled with this antibody and, in addition, labeling was found on tubular extensions often connecting peroxisomes. Antibodies to alpha-tubulin were used to locate the microtubular system. Microtubules were often found in close connection to peroxisomes, suggesting interaction between peroxisomes and the cytoskeleton. Double-labeling experiments for the 70 kDa integral peroxisomal membrane protein and alpha-tubulin demonstrated that the tubular structures connecting peroxisomes did not colocalize with microtubules. These results suggest that peroxisomes are organized in reticular structures within rat liver cells and that the structure and localization of these reticuli may be determined by their association to the microtubular network.
最近研究表明，过氧化物酶体与微管相互作用，这会影响该细胞器的结构和细胞内分布（施拉德，M.，J.K.伯克哈特，E.鲍姆加特，G.吕尔斯，H.斯普林，A.福尔克尔，H.D.法希米：微管与过氧化物酶体的相互作用。HepG2细胞中的管状和球形过氧化物酶体及其由微管活性药物诱导的改变。《欧洲细胞生物学杂志》69，24 - 35（1996年））。在本研究中，我们应用免疫学技术来研究大鼠肝细胞内过氧化物酶体的组织情况。针对与70 kDa整合型过氧化物酶体膜蛋白的403 - 417氨基酸残基相对应的十五肽制备了抗体，并在兔体内产生，然后进行亲和纯化。通过酶联免疫吸附测定（ELISA）和蛋白质印迹分析确定，该抗体对过氧化物酶体具有高度特异性。对大鼠肝脏组织切片进行免疫电子显微镜检查发现，过氧化物酶体膜被该抗体标记，此外，在通常连接过氧化物酶体的管状延伸部分也发现有标记。使用抗α - 微管蛋白抗体来定位微管系统。经常发现微管与过氧化物酶体紧密相连，这表明过氧化物酶体与细胞骨架之间存在相互作用。对70 kDa整合型过氧化物酶体膜蛋白和α - 微管蛋白进行双重标记实验表明，连接过氧化物酶体的管状结构与微管并不共定位。这些结果表明，过氧化物酶体在大鼠肝细胞内以网状结构组织，并且这些网状结构的结构和定位可能由它们与微管网络的关联所决定。
