De Souza Andrea G, MacCormack Tyson J, Wang Nan, Li Liang, Goss Greg G
Department of Chemistry, University of Alberta, Alberta, Canada.
Zebrafish. 2009 Sep;6(3):229-38. doi: 10.1089/zeb.2009.0591.
Zebrafish are an important model in vertebrate genetics, developmental biology, physiology, and toxicology. In this study, we established the first large-scale proteome profile of a teleost fish tissue using a shotgun method based on two-dimensional liquid chromatography-electrospray ionization tandem mass spectrometry. Proteome coverage was significantly improved with the application of a sequential protein solubilization method for protein fractionation and a precursor ion exclusion method for improving peptide and protein identification efficiency. Five thousand seven hundred sixteen proteins were identified with an estimated false-positive matching rate of 1.34%, and the proteome exhibited excellent coverage of important biochemical pathways relevant to the function of the gill in respiration, ion and acid-base homeostasis, and energy metabolism. Numerous established and potential biomarkers of stress, disease, and environmental contamination were also expressed in the gill. Annotation information was completely lacking for >30% of the detected proteins, highlighting the need for advancements in bioinformatics analysis techniques to complement this research. Nevertheless, the results provide important insights into the physiological function of the gill as well as its role as an environmental interface. We discuss the significance of these findings in the context of exploratory physiological and toxicological studies.
斑马鱼是脊椎动物遗传学、发育生物学、生理学和毒理学领域的重要模型。在本研究中，我们基于二维液相色谱 - 电喷雾电离串联质谱的鸟枪法，建立了首个硬骨鱼类组织的大规模蛋白质组图谱。通过应用用于蛋白质分级分离的顺序蛋白质溶解方法和用于提高肽段及蛋白质鉴定效率的前体离子排除方法，蛋白质组覆盖率得到显著提高。共鉴定出5716种蛋白质，估计假阳性匹配率为1.34%，该蛋白质组在与鳃在呼吸、离子和酸碱平衡以及能量代谢功能相关的重要生化途径方面表现出优异的覆盖率。鳃中还表达了许多已确定的和潜在的应激、疾病及环境污染生物标志物。超过30%的检测蛋白质完全缺乏注释信息，这凸显了生物信息学分析技术取得进展以补充本研究的必要性。尽管如此，这些结果为鳃的生理功能及其作为环境界面的作用提供了重要见解。我们在探索性生理学和毒理学研究的背景下讨论了这些发现的意义。
