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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。
In order to establish a two-dimensional gel electrophoresis pattern and mass spectrometry identification of fish skin proteome, the crucian carp was used as experimental animal and the protein was separated by 2-DE and MALDI-TOF-MS Proteomics of crucian carp skin. The results showed that when the amount of sample was 800μg, using 18 cm, p H3 10 IPG strips, isoelectric focusing 80 000 Vh, 214 skin protein spots were detected by 2-DE pattern, the matching rate was 88% Protein spots were identified by MS / MS as 22 proteins including myosin, actin, keratin, zinc finger protein 585B, effector protein Rab15, coiled coil protein 168, NCK1-related protein, creatine kinase, Alcoholic enzyme, pyruvate dehydrogenase, adenylate kinase isoenzyme, etc .; protein functional clustering (GO) analysis found 12 biological processes, mainly involved in metabolism, cells, organisms, immune response, biological regulation and signaling process. This study for the first time reveals the crucian carp proteomics and molecular mechanisms in order to provide scientific reference for the prevention and treatment of fish diseases.