文章摘要
张涛,宋世震,刘睿.基于nano LC-MS/MS法的黄明胶胶原蛋白鉴定研究[J].南京中医药大学学报,2020,36(1):105-109.
基于nano LC-MS/MS法的黄明胶胶原蛋白鉴定研究
Investigation on Identifying Collagen from Colla Corii Bovis Using Nano LC-MS/MS Method
  
DOI:
中文关键词: 关键词:黄明胶  胶原蛋白  鉴定  修饰  物质基础
英文关键词: Colla Corii Bovis  collagen  identification  modification  material basis
基金项目:
作者单位
张涛1,宋世震1,刘睿2,3,4 1.武汉科技大学医学院湖北 武汉 4300652.江苏省海洋药用生物资源研究与开发重点实验室江苏 南京 2100233.江苏省中药资源产业化过程协同创新中心/中药资源产业化与方剂创新药物国家地方联合工程研究中心江苏 南京 2100234.南京中医药大学药学院江苏 南京 210023 
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中文摘要:
      目的 基于shotgun混合蛋白鉴定技术,系统研究黄明胶中胶原类物质组成,探讨其胶原蛋白的修饰。方法 采用胰蛋白酶酶切黄明胶样品,纳升液相串联质谱(nano-LC Q Exactive Orbitrap MS)技术,对黄明胶酶解液进行分析获得MS/MS图谱,利用PEAKS软件对获得的MS/MS图谱搜索牛科蛋白质库,以鉴定其中蛋白质组成。结果 从2个批次的黄明胶酶解液样品中鉴定了1 378个肽段,共鉴定了30蛋白质,胶原I蛋白α1链、α2链,胶原Ⅲ蛋白α1链为黄明胶的主要物质基础,研究发现黄明胶中蛋白主要有4种修饰形式:羟基化(Hydroxylation)、脱酰胺化(Deamidation)、N-端酰胺化(Acetylation)及氧化(Oxidation)修饰。结论 黄明胶的主要物质基础源于胶原Ⅰ蛋白和胶原Ⅲ蛋白,胶原蛋白经熬制后,变性、溶出、修饰获得的产物共同构成了黄明胶的物质基础。
英文摘要:
      OBJECTIVE To investigate the proteins and peptides in Colla Corii Bovis and indicate the possible modifications in Colla Corii Bovis by Shotgun based proteomics analysis. METHODS Trypsin was used to digest two batches of Colla Corii Bovis samples, and nano-LC Q Exactive Orbitrap mass spectrometry was applied to obtain the MS/MS spectra of Colla Corii Bovis digestion samples. Then PEAKS software was used to search those MS/MS spectra against Bovine protein database to identify peptides and proteins from Colla Corii Bovis samples. RESULTS As a result, totally 30 proteins and 1 378 peptides were identified, most of the peptides were derived from Collagen Ⅰ α1 chain, Collagen Ⅰ α2 chain, and Collagen Ⅲ α1 chain. In addition, there were four modifications indicated in the present study, including Hydroxylation, Deamidation, N-term Acetylation and Oxidation. CONCLUSION The material basis of Colla Corii Bovis was mainly from Collagen Ⅰ and Collagen Ⅲ, and those components after boiled, denatured, dissolved, and modified consist of the material basis of Colla Corii Bovis.
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