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 201203,China Shanghai Institute of Pharmaceutical Industry Shanghai 201203,China.
Shanghai Institute for Food and Drug Control Shanghai 201203,China.
Zhongguo Zhong Yao Za Zhi. 2019 Jan;44(2):324-331. doi: 10.19540/j.cnki.cjcmm.20181009.012.
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.
在本研究中，以源自参环毛蚓的新鲜广地龙为研究对象。首先根据分子量通过SDS-PAGE分离广地龙的总蛋白，然后进行胶内消化。之后，通过纳升液相色谱/轨道阱融合卢莫斯高分辨率质谱（nano LC/orbitrap fusion lumos HR-MS）分析肽段。使用蛋白质组发现者软件与环节动物.fasta数据库进行比对来实现蛋白质鉴定。结果初步鉴定出386种蛋白质，包括链F、球蛋白B链、甘油醛-3-磷酸脱氢酶、纤溶蛋白等。大多数蛋白质参与细胞结构和能量代谢，纤溶蛋白和地龙激酶可能与纤溶活性有关。使用PANTHER和KEGG对代谢途径富集进行基于基因本体论的蛋白质分类。结果表明这些蛋白质与多种信号转导途径有关，包括代谢途径、中心碳代谢、氨基酸生物合成、核糖体、糖酵解、柠檬酸循环（TCA循环）等。本研究将为进一步研究广地龙中的蛋白质及其功能奠定基础。
