鲫皮肤蛋白质组双向电泳与部分蛋白点的质谱鉴定  

Preliminary proteomic analysis of the skin from crucian carp(Carassius auratus)

作　　者：胡秀彩[1] 沈晓静[2] 吕爱军[1,3] 孙敬锋[1] YEONG Yiksung 宋亚娇[3] 李莉[3] 孔祥会[3] 

机构地区：[1]天津农学院水产学院,天津市水产生态及养殖重点实验室,天津300384 [2]江苏师范大学生命科学学院,江苏徐州221116 [3]河南师范大学水产学院,河南新乡453007

出　　处：《水产学报》2016年第4期528-536,共9页Journal of Fisheries of China

基　　金：国家自然科学基金(31272692);天津市应用基础与前沿技术研究重点项目(15JCZDJC34000);河南省高校科技创新团队支持计划(15IRTSTHN018)~~

摘　　要：为了建立鱼类皮肤蛋白质组双向电泳图谱及质谱鉴定,以鲫为实验动物,采用蛋白双向电泳（2-DE）结合基质辅助激光解吸电离飞行时间质谱（MALDI-TOF/TOF-MS）技术,进行鲫皮肤的蛋白质组学研究。结果表明,当上样量为800μg,采用18 cm、p H3-10 IPG胶条,等电聚焦80 000 Vh,2-DE图谱检测为214个皮肤蛋白点,匹配率达88%;对其中31个蛋白点进行MS/MS鉴定为22种蛋白,包括肌球蛋白、肌动蛋白、角蛋白、锌指蛋白585B、效应蛋白Rab15、卷曲螺旋结构域蛋白168、NCK1相关蛋白,肌酸激酶、烯醇化酶、丙酮酸脱氢酶、腺苷酸激酶同工酶等;进行蛋白功能聚类（GO）分析发现12种生物学过程,主要参与代谢、细胞、有机体、免疫应答、生物调节和信号等过程。本研究首次初步揭示鲫皮肤蛋白质组及分子机制,以期为鱼类病害的预防和治疗提供科学参考。Fish skin plays an important role in the mucosal immune response. In order to investigate the molecular mechanisms of mucosal immune responses in fish, the Crucian carp （Carassius auratus） were used as an animal model for fish skin immune system （FSIS）, and proteomics analysis of protein expression in the skin of C. auratus was performed by using two-dimensional polyacrylamide gel electrophoresis （2-DE） combined with matrix- assisted laser desorption/ionization time-of-flight mass spectrometry （MALDI-TOF/TOF-MS） techniques. The results showed that 214 protein spots observed on 2-DE gels shared matching profiles of about 88% protein expression spots, while the 800 μg loading dose, and 18 cm pH3-10 IPG strips were used for isoelectric focusing （IEF） to 80 000 Vh. The 31 protein spots in the skin offish were further identified as tile 22 kinds of proteins by MS/MS analysis, namely myosin heavy chain, actin, keratin, zinc finger protein 585B, Rabl5 effector protein, coiled-coil domain-containing protein 168, NCKl-associated protein, and creatine kinase M-type, enolase, pyruvate dehydrogenase E1 component subunit alpha, adenylate kinase isoenzyme 1, etc. Further analysis of protein functional classification based on Gene ontology （GO） showed 12 mainly biological processes, including metabolic process, cellular process, organism process, immune response, biological regulation and signaling etc.. The three of all biological processes such as metabolic, cellular and organism accounted for 24.7%, 19.5%, and 16.9% respectively, and immune response-related proteins included transformation/transcription domain- associated protein, keratin type I cytoskeletal 10, keratin type II cytoskeletal 8, and keratin type II cytoskeletal 6A. This is the first report on proteomics analysis of expressed proteins in the skin of C. auratus, and this study will help to understand the molecular mechanisms ofmucosal immune responses at protein level in fish.

关 键 词：鲫 皮肤 蛋白质组 双向电泳 质谱 

分 类 号：Q786[生物学—分子生物学] S917.4[农业科学—水产科学]