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目的探讨肌萎缩脊髓硬化症(ALS)模型小鼠脊髓组织胶质纤维酸性蛋白(GFAP)中赖氨酸的乙酰化修饰。方法对4个月左右的ALS模型小鼠进行麻醉处死,提取小鼠脊髓组织总蛋白,通过GFAP抗体和乙酰化抗体,运用免疫沉淀和免疫蛋白印迹技术鉴定GFAP蛋白的表达与GFAP蛋白赖氨酸的乙酰化修饰,通过液相二级质谱连用(LC-MS/MS)检测ALS模型小鼠脊髓组织中GFAP赖氨酸的乙酰化修饰位点。结果免疫沉淀和免疫蛋白印迹方法发现ALS模型小鼠脊髓组织中GFAP蛋白存在赖氨酸乙酰化修饰;LC-MS/MS发现GFAP的394位和402位赖氨酸位点发生乙酰化修饰。结论 ALS小鼠模型脊髓组织中,GFAP的赖氨酸位点能被乙酰化修饰,该修饰可能参与了ALS的发生。
Objective To investigate the acetylation of lysine in glial fibrillary acidic protein (GFAP) of amyotrophic lateral sclerosis (ALS) model mice. Methods ALS model mice were anesthetized and sacrificed at 4 months. The total protein of spinal cord tissue of mice was extracted. The expression of GFAP protein and GFAP protein lysine were determined by immunoprecipitation and immunoblotting using GFAP antibody and acetylation antibody. The acetylated sites of GFAP lysine in the spinal cord of ALS model mice were detected by liquid chromatography-mass spectrometry (LC-MS / MS). Results The results of immunoprecipitation and immunoblotting showed that there was lysine acetylation of GFAP protein in spinal cord of mice with ALS model. The acetylation of GFAP at positions 394 and 402 was detected by LC-MS / MS. Conclusion ALS mouse model of spinal cord tissue, GFAP lysine sites can be acetylated, the modification may be involved in the occurrence of ALS.