Departamento de Bioquímica y Biología Molecular IV, Facultad de Veterinaria, Universidad Complutense de Madrid, 28040 Madrid, Spain.
J Proteome Res. 2010 Dec 3;9(12):6392-404. doi: 10.1021/pr100683k. Epub 2010 Nov 5.
We report the novel use of proteomics to investigate protein variation among populations of the European hake (Merluccius merluccius). The liver and brain extracts of 18 hake (N = 36) captured in the Mediterranean Sea, Cantabrian Sea, and Atlantic Ocean were examined by 2D/DIGE and mass spectrometry. Significant differences in protein expression among populations were revealed by 84 spots obtained in the gels for the liver and 145 spots for the brain. Population groups of samples were defined by multivariate analysis (PCA and hierarchical clustering). According to protein expression levels and the functions of the 55 candidate protein spots identified, which showed significant expression differences, highest population discrimination was rendered by brain proteins involved in cell signaling and metabolism/energy and by liver proteins involved in protein fate. Finally, we present a statistically robust framework to accurately classify individuals according to their population of origin. Thus, purposely identified protein isoforms were found to be competent at discriminating populations. These results suggest the possibility of identifying protein biomarkers related to environmental changes in a nonmodel species such as the hake and pave the way for more extensive research on protein variation among populations of marine fishes.
我们报告了蛋白质组学在研究欧洲无须鳕(Merluccius merluccius)种群间蛋白质变异中的新应用。通过二维电泳/DIGE 和质谱法分析了地中海、坎塔布连海和大西洋捕获的 18 条无须鳕(N=36)的肝脏和大脑提取物。肝脏凝胶中获得的 84 个斑点和大脑中获得的 145 个斑点揭示了种群间蛋白质表达的显著差异。通过多元分析(PCA 和层次聚类)定义了样品的种群组。根据蛋白质表达水平和鉴定的 55 个候选蛋白斑点的功能,这些斑点显示出显著的表达差异,脑蛋白中与细胞信号转导和代谢/能量有关的蛋白以及肝蛋白中与蛋白质命运有关的蛋白对种群的区分度最高。最后,我们提出了一个统计上稳健的框架,可以根据个体的种群来源准确地对其进行分类。因此,发现特意鉴定的蛋白同工型能够区分种群。这些结果表明,在无须鳕等非模式物种中识别与环境变化相关的蛋白质生物标志物是有可能的,并为海洋鱼类种群间蛋白质变异的更广泛研究铺平了道路。