Fernández-Boo S, Chicano-Gálvez E, Alhama J, Barea J L, Villalba A, Cao A
Centro de Investigacións Mariñas, Consellería do Medio Rural e do Mar, Xunta de Galicia, Aptdo. 13, 36620 Vilanova de Arousa, Spain.
Department Biochemistry & Molecular Biology, University of Córdoba, Córdoba, Spain; Instituto Maimónides de Investigación Biomédica de Córdoba (IMIBIC)/Hospital Universitario Reina Sofía/Universidad de Córdoba, Spain.
J Invertebr Pathol. 2014 May;118:47-58. doi: 10.1016/j.jip.2014.02.011. Epub 2014 Mar 5.
The genus Perkinsus includes protozoan parasites of a wide range of marine molluscs worldwide, some of which have been responsible for heavy mollusc mortalities and dramatic economic losses. This study was performed with the aim of increasing the knowledge of Perkinsus spp. proteome. Proteins extracted from in vitro cultured cells of three species of this genus, P. marinus, P. olseni and P. chesapeaki, were analysed using 2D electrophoresis. Four gels from each species were produced. Qualitative and quantitative comparisons among gels were performed with Proteamweaver software. Cluster analysis grouped the four gels of each Perkinsus sp.; furthermore, P. marinus and P. olseni gels were grouped in a cluster different from P. chesapeaki. Around 2000 spots of each species were considered, from which 213 spots were common to the 3 species; P. chesapeaki and P. marinus shared 310 spots, P. chesapeaki and P. olseni shared 315 spots and P. marinus and P. olseni shared 242 spots. A number of spots were exclusive of each Perkinsus species: 1161 spots were exclusive of P. chesapeaki, 1124 of P. olseni and 895 of P. marinus. A total of 84 spots, including common and species-specific ones, were excised from the gels and analysed using MALDI-TOF and nESI-IT (MS/MS) techniques. Forty-two spots were successfully sequenced, from which 28 were annotated, most of them clustered into electron transport, oxidative stress and detoxification, protein synthesis, carbohydrate metabolism, signal transduction, metabolic process and proteolysis.
派金虫属包括世界各地多种海洋软体动物的原生动物寄生虫，其中一些已导致大量软体动物死亡并造成巨大经济损失。本研究旨在增加对派金虫属蛋白质组的了解。使用二维电泳分析了从该属的三个物种，即马里派金虫、奥尔森派金虫和切萨皮克派金虫的体外培养细胞中提取的蛋白质。每个物种制备了四张凝胶。使用Proteamweaver软件对凝胶进行定性和定量比较。聚类分析将每个派金虫属物种的四张凝胶分组；此外，马里派金虫和奥尔森派金虫的凝胶被归为与切萨皮克派金虫不同的一个聚类。每个物种约2000个斑点被纳入考虑，其中213个斑点是三个物种共有的；切萨皮克派金虫和马里派金虫共有310个斑点，切萨皮克派金虫和奥尔森派金虫共有315个斑点，马里派金虫和奥尔森派金虫共有242个斑点。每个派金虫属物种都有一些独特的斑点：1161个斑点是切萨皮克派金虫独有的，1124个是奥尔森派金虫独有的，895个是马里派金虫独有的。总共从凝胶中切下84个斑点，包括共有斑点和物种特异性斑点，并使用基质辅助激光解吸电离飞行时间质谱和纳升电喷雾离子阱串联质谱（MS/MS）技术进行分析。42个斑点成功测序，其中28个得到注释，它们大多聚集成电子传递、氧化应激与解毒、蛋白质合成、碳水化合物代谢、信号转导、代谢过程和蛋白水解等类别。
