有毒甲藻Alexandrium catenella DH01的蛋白质组学研究:使用荧光标记的方法检测和鉴定细胞表面蛋白

lexandrium catenella DH01 is a toxic dinoflagellate species which is not only able to produce paralytic shellfish toxins(PSTs) but also cause harmful algal blooms (HABs) in the coast of China. In this study, we present a new protocol for specific labeling and detection of cell surface proteins (CSPs) of A. catenella DH01 cells using CyDye DIGE Fluor minimal dyes, and have identified CSPs using a combining approach of two dimensional gel electrophoresis (2-DE) and MALDI-TOF-TOF mass spectrometry. The results showed that the fluorescent cyanine dye Cy3 could specific combine with CSPs of A. catenella DH01 cells with minimal labeling of intracellular proteins. Trizol method was the best to extract CSPs with respect to protein spot number and resolution among the three protein extraction methods. A total of 41 CSPs were separated and identified from A. catenella DH01 in 2-D gel, among of them, 14 proteins gave the positive identification of protein orthologues in the protein database by MALDI TOF-TOF MS analysis. This work represents the first attempt to investigate CSPs of A. catenella using CyDye DIGE fluors dyeing method, which provides a potential important tool for future comprehensive characterization of CSPs and elucidation of physiological functions of CSPs in dinoflagellatess.