蒙古黄芪和膜荚黄芪水溶性蛋白表达

目的:蒙古黄芪和膜荚黄芪都为药用黄芪,二者亲缘关系近,成分相似,药用价值尚未区分。该研究旨在建立蒙古黄芪和膜荚黄芪的水溶性蛋白表达谱,了解两种黄芪之间存在的差别。方法:样品采用水超声提取和丙酮沉淀的方法得到黄芪水溶性蛋白成分,质谱级胰酶酶解后的肽段经nano ESI-LC-MS/MS分析,采用Proteome Discoverer 1. 4软件比对豆科蛋白数据库鉴定蛋白,再使用非标记定量软件SIEVE分析蒙古黄芪与膜荚黄芪水溶性蛋白质表达的差异。最后运用生物信息学方法分析2种黄芪共有蛋白的分类、分子功能、参与的生物过程和信号通路。结果:蒙古黄芪鉴定特异性蛋白有920个,膜荚黄芪鉴定的特异性蛋白有717个,2种黄芪中共同表达的蛋白有472个,其中二者的差异表达蛋白有21个,如PR-10蛋白,NDK-1蛋白,谷蛋白A2,磷脂酶D等蛋白在两种黄芪中差异表达。蒙古黄芪高表达蛋白14个,膜荚黄芪高表达蛋白7个。结论:蒙古黄芪和膜荚黄芪的水溶性蛋白表达谱存在显著差异,特异性蛋白、差异表达蛋白和共同表达的蛋白可为今后两种黄芪的鉴别、药理作用机制的研究和寻找作用靶点提供参考。

Ater-soluble Protein Profiles of Astragalus membranaceus var. mongholicus and A. membranaceus

Objective: Astragalus membranaceus var. mongholicus and A. membranaceus are medicinal Astragalus,which are closely related and similar in composition,but with unclear medicinal value. Water-soluble protein profiles for A. membranaceus var. mongholicus and A. membranaceus were established to explore the differences between the two kinds of Astragalus Radix. Method: The water-soluble protein components were obtained through water ultrasonic extraction and acetone precipitation. After digested with trypsin,the obtained peptides were analyzed by nano ESI-LC-MS/MS method. Proteome Discovery 1.4 software was used to identify the proteins by comparing with the legume protein database,and the different expression water-soluble proteins were analyzed by the label-free quantitative software SIEVE. Finally,relevant information for common expression proteins,including classification,molecular function,involved biological process and signaling pathway, were analyzed by bioinformatics. Result: There were 920 and 717 specific proteins identified for A. membranaceus var. mongholicus and A. membranaceus,respectively. Totally 472 proteins were found to be co-expressed,in which 21 were differentially expressed,such as PR-10 protein,NDK-1 protein,glutelin A₂,and phospholipase D. There were 14 highly expressed proteins in A. membranaceus var. mongholicus and 7 highly expressed proteins in A. membranaceus. Conclusion: There are significant differences in water-soluble protein profiles for two kinds of Astragalus Radix. Specific proteins, differentially expressed proteins and common expressed proteins can provide references for the identification of A.membranaceus var. mongholicus and A. membranaceus. It also can be used to define pharmacological mechanisms and search for drug action targets.