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目的论证重症肌无力(MG)患者骨骼肌中特异性减少的25000蛋白为碳酸酐酶Ⅲ(CAⅢ)分子。方法用双向电泳结合免疫Western blot分析25000蛋白抗体和CAⅢ抗体有免疫反应的蛋白质分子的物化特征,分别用免疫Dot blot与免疫Western blot观察两种抗体对纯化的25000蛋白及骨骼肌蛋白匀浆的竞争结合,用免疫Western blot分析健康人与MG患者骨骼肌CAⅢ的蛋白表达。结果双向电泳结合免疫Western blot显示,25000蛋白抗体和CAⅢ抗体识别的蛋白质具有相同的相对分子质量和等电点;免疫Dot blot与免疫Western blot的竞争结合证实25000蛋白与CAⅢ为同一物质;免疫Western blot表明,用25000蛋白抗体与CAⅢ抗体检测健康人与MG患者骨骼肌25000蛋白表达的结果亦几乎是相同的。结论MG患者骨骼肌特异减少的25000蛋白分子就是CAⅢ,这为进一步深入研究CAⅢ与MG的发病奠定了基础。
Objective To demonstrate that the 25000 protein that is specifically reduced in skeletal muscle of patients with myasthenia gravis (MG) is a carbonic anhydrase III (CA III) molecule. Methods Two-dimensional electrophoresis combined with Western blot analysis of 25000 protein antibody and CA Ⅲ antibody immunoreactive protein molecular physical and chemical characteristics, respectively, by immunoblot blot and immunoblot Western blot were observed two antibodies to purified 25000 protein and skeletal muscle protein homogenate Competition and binding, Western blot analysis of healthy people and MG patients with skeletal muscle CA Ⅲ protein expression. Results Two-dimensional electrophoresis combined with immunoprecipitation Western blot showed that the proteins identified by 25000 and CAⅢ antibodies had the same relative molecular mass and isoelectric point. The competitive immunoblotting of immunoblot blot and Western blot confirmed that 25000 protein was the same as CAⅢ, The results of blot showed that the results of 25000 protein expression in skeletal muscle of healthy and MG patients by using 25000 protein antibody and CA Ⅲ antibody were also almost the same. Conclusion The 25 000 protein molecule with specific reduction of skeletal muscle in patients with MG is CA Ⅲ, which lays the foundation for further study of the pathogenesis of CA Ⅲ and MG.