中国中药杂志

2019, v.44(02) 324-331

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基于纳升高效液相色谱-四极杆-线性离子阱-静电场轨道阱高分辨质谱技术研究广地龙中的蛋白质
Study on proteins in Guangdilong by nano LC/orbitrap fusion lumos HR-MS

董洪霜;张静娴;胡青;王艳春;孙健;张甦;于泓;冯睿;毛秀红;季申;
DONG Hong-shuang;ZHANG Jing-xian;HU Qing;WANG Yan-chun;SUN Jian;ZHANG Su;YU Hong;FENG Rui;MAO Xiu-hong;JI Shen;Shanghai Institute for Food and Drug Control;Shanghai Institute of Pharmaceutical Industry;Shanghai Kangqiao Chinese Medicine Tablet Co.,Ltd.;

摘要(Abstract):

该文以新鲜广地龙(来源钜蚓科动物参环毛蚓Pheretima aspergillum)为研究对象,采用SDS-PAGE法对广地龙总蛋白进行分离,并进行胶内酶切,利用纳升高效液相色谱-四极杆-线性离子阱-静电场轨道阱高分辨质谱技术研究广地龙中的蛋白质。采用Proteome Discoverer软件,检索环节动物门蛋白数据库(Annelida. fasta)鉴定蛋白。结果共鉴定到386个蛋白质,包括珠蛋白、甘油醛-3-磷酸脱氢酶、纤溶活性蛋白等,大多数蛋白质与细胞结构、能量供给有关,纤溶活性蛋白、胍乙基磷酸丝氨酸蛋白酶可能与抗凝血或溶栓活性有关。利用PANTHER对蛋白进行基因本体功能分类,KEGG进行代谢通路富集分析,发现涉及多种信号转导通路,包括代谢途径、碳代谢通路,氨基酸的生物合成通路,糖酵解/糖异生通路,柠檬酸循环等54种。该研究为阐明广地龙的蛋白物质基础以及其功能研究提供了实验依据。
In the present study,fresh Guangdilong( GD),originating from Pheretima aspergillum,was taken as the object. The total proteins from GD were firstly separated by SDS-PAGE according to their molecular weights and in-gel digestion was then performed.After that,the peptides were analyzed by nano LC/orbitrap fusion lumos high resolution mass spectrometry( nano LC/orbitrap fusion lumos HR-MS). Protein identification was implemented by comparison with Annelida. fasta database using Proteome Discoverer software.As a result,386 proteins were tentatively identified,including chain F,globin B chain,glyceraldehyde-3-phosphate dehydrogenase,fibrinolytic protein,and so on. Most of the proteins took part in cell structure and energy metabolism,and fibrinolytic protein and lombricine kinase might be related to fibrinolytic activity. Protein classification based on gene ontology was carried out using PANTHER and KEGG for metabolic pathway enrichment. The results indicated that these proteins were related to diverse signal transduction pathways,including metabolic pathways,central carbon metabolism,biosynthesis of amino acids,ribosome,glycolysis,citrate cycle( TCA cycle),and so on. This study would lay the foundation for the further research on the proteins in GD and also their functions.

关键词(KeyWords): 广地龙;蛋白质;SDS-PAGE;orbitrap fusion lumos
Guangdilong;proteome;SDS-PAGE;orbitrap fusion lumos

Abstract:

Keywords:

基金项目(Foundation): 上海市科委课题项目(13DZ1970402);; 上海市中药和保健食品品质与安全检测专业技术服务平台项目(18DZ2292200)

作者(Author): 董洪霜;张静娴;胡青;王艳春;孙健;张甦;于泓;冯睿;毛秀红;季申;
DONG Hong-shuang;ZHANG Jing-xian;HU Qing;WANG Yan-chun;SUN Jian;ZHANG Su;YU Hong;FENG Rui;MAO Xiu-hong;JI Shen;Shanghai Institute for Food and Drug Control;Shanghai Institute of Pharmaceutical Industry;Shanghai Kangqiao Chinese Medicine Tablet Co.,Ltd.;

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DOI: 10.19540/j.cnki.cjcmm.20181009.012

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