基于nano LC-LTQ/Orbitrap MS分析山羊角中蛋白质多肽类物质

Analysis and Identification of Proteins and Peptides from Goat Horn (Caprae Hircus Cornu) Using nano LC-LTQ/Orbitrap MS

  • 摘要: 基于shotgun混合蛋白质鉴定思路,研究山羊角水提液中蛋白质类、肽类物质组成。采用超高分辨的线性离子阱-静电场轨道阱质谱(LTQ/Orbitrap MS),从山羊角水提液中大于3 ku部位鉴定出52个蛋白质,从小于3 ku部位鉴定出1288个肽段,这些肽段源于46个蛋白质。鉴定出的蛋白质主要为角蛋白、胶原蛋白、角蛋白相关蛋白等结构蛋白。根据每个氨基酸出现的频次,将蛋白质上的肽段标记,绘制热图(heat map),据此推测山羊角水提液中多肽类物质形成的过程为:从蛋白质的热区(hot regions)中经过降解、脱落、溶出,形成水提液中的各种肽段。该方法能够快速、高效地分析鉴定山羊角水提液中蛋白质肽类物质组成,可用于角类动物药蛋白质类物质组成与分析,也可为中药动物药的系统研究提供借鉴与方法参考。

     

    Abstract: The objective of the present study was to study the proteins and peptides of goat horn (Caprae Hircus Cornu, GH) based on the shotgun strategy. By using LTQ/Orbitrap MS, proteins and peptides were identified from GH aqueous extract. In total, 52 proteins were identified from GH fraction with the molecular weight larger than 3 ku, and 1288 peptides were identified from GH fraction with the molecular weight lower than 3 ku, and these peptides were hydrolyzed from 46 proteins. The identified proteins were keratin, collagen, keratin-associated protein, etc. It can be speculated that the particular “hot regions” in the amino acid sequence which were easier to proteolysis, drop, dissolve and generate a number of peptides. According to the present strategy, proteins and peptides derived from GH aqueous extract can be rapid and efficient identified. The strategy can be applied in analysis and identification of proteins in animal horn derived TCMs, and provide a reference for investigate the material basis of animal horn derived TCMs.

     

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