人肌肉乙酰胆碱受体的初步纯化及其生物学鉴定

Preliminary Purification and Identification of Acetylcholine Receptor Extracted from Human Bone Muscle

目的 :从人肌肉中提取具有免疫活性的乙酰胆碱受体蛋白 ,为复制MG动物模型提供抗原。方法 :新鲜股四头肌制成匀浆后 ,采用TritonX - 1 0 0提取 ,以SephadexG - 1 50柱凝胶层析 ,初步纯化 ;然后采用紫外分光光度法及SDS -聚丙烯酰胺凝胶电泳 ,检测蛋白的含量和纯度 ;应用主动免疫的方法 ,检测AchR的免疫学特性。结果 :此纯化物的蛋白含量为 3 .2mg/ml;SDS -聚丙烯酰胺凝胶电泳 ,呈一条 44kD的主要蛋白带 ;此纯化物成功地复制出了MG动物模型。结论 :该方法提取的乙酰胆碱受体蛋白 ,具有免疫活性 。

Objective To extract and purify acetylcholine receptor (AchR) from human bone muscle. Methods Using Triton X-100 extraction,ultra centrifation and sephadex G-150 filtration, AchR was extracted and preliminarily purified from human bone muscle,and identified by ultraviolet light absorbance,SDS- polyacrylamide gel electrophoresis (SDS-PAGE) and initiative immunity. Results The content of the protein was 3.2 mg/ml. One main band of 44 kD peptide was on SDS-polyacrylamide gel. And the AchR protein induced experimental autoimmune myasthenia gravis (EAMG) successfully. Conclusion The AchR protein extracted by the method had the immunocompetence,which could serve as the antigen to induce EAMG.