Evidence for an immunological and functional relationship between superoxide dismutase and a high molecular weight osteoclast plasma membrane glycoprotein.
Evidence for an immunological and functional relationship between superoxide dismutase and a high molecular weight osteoclast plasma membrane glycoprotein.
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超氧化物歧化酶和高分子量破骨细胞质膜糖蛋白之间免疫学和功能关系的证据。
DOI：
10.1002/jcb.240460408
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发表时间：
1991
影响因子：
4
通讯作者：
Osdoby,P
中科院分区：
文献类型：
--
作者：
Oursler,MJ;Collin-Osdoby,P;Li,L;Schmitt,E;Osdoby,P
大型多核破骨细胞是负责骨分解的主要细胞，据报道可产生高水平的超氧化物，这可能有助于骨吸收过程（Key 等人：J Bone and Mineral Res 4 [suppl. 1]：S206，1989）。破骨细胞还具有高水平的超氧化物歧化酶，这是一种能够将有毒超氧化物转化为毒性较小的 H 2 O 2 的保护酶（Fridovich: J Biol Chem 264: 7761–7764, 1989）。锰和/或铁超氧化物歧化酶的氨基酸序列具有保守区，该保守区表现出与从与单克隆抗体121F反应的高分子量破骨细胞表面标记糖蛋白获得的片段基本同源性。在本报告中，提供的证据证实了 121F 单克隆抗体识别的破骨细胞膜抗原和超氧化物歧化酶之间的免疫学、生化和功能相似性。蛋白质印迹和免疫沉淀研究表明，针对免疫亲和纯化抗原产生的单特异性多克隆抗体与超氧化物歧化酶发生交叉反应。在破骨细胞质膜制剂中已检测到抗原和高分子量超氧化物歧化酶活性。在抗原耗竭研究和探测破骨细胞和密切相关的骨髓源性巨细胞的蛋白质印迹中，发现超氧化物歧化酶活性和膜抗原的水平相关。此外，凝胶电泳和蛋白质印迹显示，通过电泳酶谱分析鉴定的破骨细胞超氧化物歧化酶活性区域含有高分子量抗原，或当这些在非变性电泳之前预混合时，抗原与121F单克隆抗体的复合物。据认为破骨细胞质膜具有高分子量超氧化物歧化酶活性。此外，这种活性似乎与 121F 单克隆抗体识别的破骨细胞抗原有关。
Large multinucleated osteoclasts are the major cells responsible for bone breakdown and have been reported to produce high levels of superoxides which may contribute to the process of bone resorption (Key et al.: J Bone and Mineral Res 4 [suppl. 1]: S206, 1989). Osteoclasts also possess high levels of superoxide dismutase, a protective enzyme capable of converting toxic superoxides to less toxic H 2 O 2 (Fridovich: J Biol Chem 264: 7761–7764, 1989). The amino acid sequence of manganese and/or iron superoxide dismutase has a conserved region which exhibits substantial homology with a fragment obtained from a high molecular weight osteoclast surface marker glycoprotein which is reactive with monoclonal antibody 121F. In this report, evidence is presented substantiating immunological, biochemical, and functional similarities between the osteoclast membrane antigen recognized by the 121F monoclonal antibody and superoxide dismutase. Western blot and immunoprecipitation studies show that a monospecific polyclonal antibody generated against immunoaffinity purified antigen is cross-reactive with superoxide dismutase. Both the antigen and a high molecular weight superoxide dismutase activity have been detected in osteoclast plasma membrane preparations. The levels of superoxide dismutase activity and the membrane antigen have been found to correlate in antigen depletion studies and in western blots probing osteoclasts and closely related marrow-derived giant cells. Moreover, regions of osteoclast superoxide dismutase activity identified by electrophoretic zymogram analysis have been shown by gel electrophoresis and western blots to contain the high molecular weight antigen, or complexes of the antigen with the 121F monoclonal antibody when these were premixed prior to nondenaturing electrophoresis. It is proposed that the osteoclast plasma membrane possesses a high molecular weight superoxide dismutase activity. Furthermore, it appears that this activity is associated with the osteoclast antigen recognized by the 121F monoclonal antibody.