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海螵蛸蛋白质、肽类物质基础研究

刘睿1,2,3 魏爽1,2,3 王欣之1,2,3 程建明1,2,3 吴皓1,2,3

(1.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 210023)
(2.南京中医药大学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 210023)
(3.南京中医药大学药学院, 江苏南京 210023)

【摘要】基于shotgun混合蛋白鉴定技术,系统研究海洋中药海螵蛸蛋白质类、肽类物质组成。以酸化甲醇提取海螵蛸粗多肽,采用强阳离子交换树脂(SCX)进一步富集其中多肽类成分;提取海螵蛸粗蛋白,通过胰蛋白酶(trypsin)酶解制备蛋白酶解液。采用nano-LC Q Exactive Orbitrap质谱仪分析鉴定海螵蛸蛋白质、肽类成分。基于软体动物门Mollusca蛋白质数据库比对分析,从海螵蛸中共鉴定出16个蛋白质类成分,168个多肽类成分,从头测序(De novosequence)方法鉴定了328个多肽类成分。蛋白质类成分主要为血蓝蛋白(hemocyanin)、烯醇化酶(enolase)、肌球蛋白(myosin)、肌动蛋白(actin)、钙调蛋白(calmodulin)等;多肽类成分主要来源于肌动蛋白、组蛋白(histone)、微管蛋白(tubulin)等。这些蛋白质、肽类成分构成了海洋中药海螵蛸的重要物质基础。该文为海螵蛸的功效物质基础系统研究及反映传统功效的活性成分的发现提供理论依据与研究基础。

【关键词】 海洋中药;海螵蛸;蛋白质;多肽;物质基础;

【DOI】

【基金资助】 江苏省高校“青蓝工程”优秀青年骨干教师项目(2016年); 江苏省“333工程”第三层次(2016年); 江苏省海洋科技创新专项(HY2017-7); 药用南珠标准化养殖与综合利用产业链协同创新项目(Bhsfs002); 江苏省高等学校自然科学研究重大项目(17KJA360005);

Proteomics and peptidomics analysis of Sepiae Endoconcha

LIU Rui1,2,3 WEI Shuang1,2,3 WANG Xin-zhi1,2,3 CHENG Jian-ming1,2,3 WU Hao1,2,3

(1.Jiangsu Key Laboratory of Research and Development in Marine Bio-resource Pharmaceutics, Nanjing, Jiangsu, China 210023)
(2.Jiangsu Collaborative Innovation Center of Chinese Medicinal Resources Industrialization, National and Local Collaborative Engineering Center of Chinese Medicinal Resources Industrialization and Formulae Innovative Medicine, Nanjing University of Chinese Medicine, Nanjing, Jiangsu, China 210023)
(3.School of Pharmacy, Nanjing University of Chinese Medicine, Nanjing, Jiangsu, China 210023)

【Abstract】Shotgun-based proteomics and peptidomics analyses were used to investigate the proteins and peptides in marine Chinese medicinal Sepiae Endoconcha (cuttlebone). Peptides were extracted from Sepiae Endoconcha by acidified methanol, and then strong cation exchange (SCX) resin was used to enrich those peptides. Also, proteins from Sepiae Endoconcha were extracted and digested by trypsin. Nano-LC Q Exactive Orbitrap mass spectrometry was used to analyze proteins and peptides from Sepiae Endoconcha. As a result, a total of 16 proteins and 168 peptides were identified by protein database search, and 328 peptides were identified by De novo sequencing. The identified proteins were hemocyanin, enolase, myosin, actin, calmodulin, etc., and the identified peptides were derived from actin, histone, and tubulin. All these proteins and peptides were important components in Sepiae Endoconcha, which would provide important theoretical and research basis for marine Chinese medicinal Sepiae Endoconcha investigations.

【Keywords】 marine Chinese medicinal; Sepiae Endoconcha (cuttlebone); protein; peptide; material basis;

【DOI】

【Funds】 Excellent Young Teachers (Qinglan) Project of Universities in Jiangsu Province (2016); The Third Level of “333 Project” in Jiangsu Province (2016); Special Project on Marine Science and Technology Innovation in Jiangsu Province (HY2017-7); Collaborative Innovation Project of Industry Chain of Standardized Breeding and Comprehensive Utilization of Medicinal South Pearl (Bhsfs002); Major Natural Science Research Project of Higher Education Colleges in Jiangsu (17KJA360005);

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This Article

ISSN:1001-5302

CN:11-2272/R

Vol 45, No. 16, Pages 3883-3889

August 2020

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Article Outline

Abstract

  • 1 Materials
  • 2 Methods
  • 3 Results
  • 4 Discussion
  • References