纳升级反相液相色谱串联质谱法分析海马蛋白质组

目的：为了全面了解海龙的蛋白质成分，对其蛋白表达谱进行蛋白质组学分析。方法：应用三氯乙酸/丙酮沉淀法提取海马药材的蛋白质，经聚丙烯酰胺凝胶电泳分离分析，将聚丙烯酰胺凝胶电泳条带切下进行胶内酶解处理后，应用纳升级反相液相色谱串联质谱（nanoRPLC-MS/MS）法系统分析海马提取蛋白质胶内酶解产物，并进行数据库检索鉴定蛋白质。结果：共鉴定到海马蛋白192种，等电点范围为4.06~12.18，相对分子质量范围为3.06×10³~3.52×10⁶，多肽10 680个；通过生物信息学方法分析了海马提取蛋白质的分子功能。得到海马提取物中的蛋白质具有结合、酶催化、肌动、ATP酶等功能活性，其中结合活性的蛋白质占的比例较大。结论：本研究建立了海马的蛋白质表达谱，可为海马药材中蛋白质的深入研究奠定基础，对海马药材有效成分的分析和海马药材鉴别具有重要意义。

Objective:To fully understand the protein composition of Hippocampus,the protein expression profiles were analyzed by proteomics in this study.Methods:Trichloroacetic acid/acetone precipitation method was used to extract proteins from dried Hippocampus and the proteins were separated by SDS-PAGE.The protein bands were cut from the gel and digested with trypsin to generate peptide mixture.The peptide fragments were then analyzed using nanoflow reversed phase liquid chromatography-tandem mass spectrometry(nanoRPLC-MS/MS)method.The protein was identified with database searching.Results:A total of 192 proteins and 10 680 peptides were identified.The proteins relative molecular mass values ranged from 3.06×10³-3.52×10⁶ and pI values ranged from 4.06 to 12.18.In order to explore possible molecular functions of the identified proteins,bio-information analysis was conducted on the raw data using the DAVID Bioinformatics Resources 6.8 Gene Ontology(GO) analysis tool.The function of proteins included binding activity,enzyme catalytic activity,motor activity and antioxidant activity,where binding activity was the major molecular functions involved in the energy metabolism. Conclusion:The study has established the protein expression profile of Hippocampus,which can set up a foundation for further research on proteins from Hippocampus,and has significant impact on the analysis of its effective components and the identification of Hippocampus.