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。
Shotgun based proteomics and peptidomics analysis were used to investigate the proteins and peptides in marine traditional Chinese medicine(TCM)Sepiae Endoconcha(cuttlebone).Peptides were extracted from cuttlebone by acidified methanol,and then strong cation exchange(SCX)resin was used to enrich those peptides.Also,proteins from cuttlebone were extracted and digested by trypsin.nano-LC Q Exactive Orbitrap mass spectrometry was used to analyze proteins and peptides from cuttlebone.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 cuttlebone,which would provide important theoretical and research basis for marine TCM cuttlebone investigations.
作者
刘睿
魏爽
王欣之
程建明
吴皓
LIU Rui;WEI Shuang;WANG Xin-zhi;CHENG Jian-ming;WU Hao(Jiangsu Key Laboratory of Research and Development in Marine Bio-resource Pharmaceutics,Nanjing 210023,China;Jiangsu Collaborative lnnowation Center of Chinese Medicinal Resourcs lndustrialization,National and Local CollaborativeEngineering Center of Chinese Medicinal Resources lndustrialization and Formulae Innovative Medicine,Nanjing University of Chinese Medicine,Nanjing 210023,China;School of Pharmacy,Nanjing Unizersity of Chinese Medicine,Narjing 210023,Chiraa)
出处
《中国中药杂志》
CAS
CSCD
北大核心
2020年第16期3883-3889,共7页
China Journal of Chinese Materia Medica
基金
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)。
关键词
海洋中药
海螵蛸
蛋白质
多肽
物质基础
marine traditional Chinese medicine
Sepiae Endoconcha(cuttlebone)
protein
peptide
material basis
