尿蛋白电泳分析和质谱鉴定  

Electrophoretic analysis and mass spectrometric identification of urinary proteins

作　　者：王颖[1] 黄勇[1] 张小容[1] 罗高兴[1] 贺伟峰[1] 吴军[1] 

机构地区：[1]第三军医大学西南医院全军烧伤研究所/创伤烧伤及复合伤国家重点实验室/疾病蛋白组学重庆市市级重点实验室,重庆400038

出　　处：《现代医药卫生》2016年第12期1803-1805,1809,共4页Journal of Modern Medicine & Health

摘　　要：目的探究尿蛋白的最佳沉淀方法 ,且对常规损失蛋白进行鉴定分析。方法采用有机溶剂沉淀法,即用三氯乙酸/90%丙酮法、三氯乙酸/100%丙酮法和90%丙酮法分别沉淀尿液中蛋白质,对90%丙酮法所得到沉淀的可溶部分和不溶部分进行十二烷基硫酸钠-聚丙烯酰胺分离和质谱鉴定。结果 90%丙酮沉淀法获得的尿蛋白量相对较高。将90%丙酮法沉淀的尿蛋白不溶部分和可溶部分进行质谱鉴定,可溶部分得到53种蛋白,不溶部分得到49种蛋白,且不溶部分特有的蛋白包括免疫球蛋白κ轻链基因1-5、谷氨酰氨肽酶、突变体1、芳基硫酸酯酶A前体等14种。结论尿液沉淀的不溶部分中特有蛋白的发现有助于尿液蛋白质组的全谱研究,对临床特异蛋白的检测具有重要意义。Objective To explore the best urine protein precipitation method, and to identify the routine loss protein.Methods The organic solvent precipitation method,i.e.,TCA/90% acetone,TCA/100% acetone and 90% acetone,was used to precipitate urinary proteins,and the obtained soluble and insoluble proteins by the 90% acetone method were separated and iden-tified by SDS-PAGE and mass spectrum. Results The urinary proteins amounts obtained by 90% acetone precipitation were relatively higher. The soluble and insoluble parts obtained by 90% acetone precipitation were identified by mass spectrometry. Fifty-three proteins were obtained from the soluble part and 49 proteins from insoluble part,moreover the specific proteins in insoluble part included immunoglobulin kappa light chain1-5（IGKV1-5）,glutamyl aminopeptidase（ENPEP）,PROM1,arylsulfatase A（ARSA）precursor,etc. Conclusion The discovery of these proteins in insoluble parts of urine precipitation may contribute to the whole spectrum study of urinary proteome,and has an important clinical significance to the specific protein detection.

关 键 词：蛋白尿/诊断 沉淀 电泳 聚丙烯酰氨凝胶 蛋白质组学 质谱分析法 

分 类 号：R692.5[医药卫生—泌尿科学]