Proteomics of a toxic dinoflagellate Alexandrium catenella DH01: Detection and identification of cell surface proteins using fluorescent labeling

Abstract

Alexandrium catenella DH01 is a toxic dinoflagellate species that is able to not only produce paralytic shellfish toxins, but also cause harmful algal blooms along the coast of China. In this study, we presented a new protocol for specific labeling and detection of the cell surface proteins (CSPs) of A. catenella DH01 cells using CyDye difference gel electrophoresis (DIGE) fluor minimal dyes. CSPs were identified using two-dimensional gel electrophoresis (2-DE) and MALDI TOF-TOF mass spectrometry (MS). The results showed that the fluorescent cyanine dye Cy3 could specifically label the CSPs of A. catenella DH01, with minimal labeling of intracellular proteins. Among three protein extraction methods evaluated, the Trizol method was the most efficient to extract CSPs with respect to protein spot number and resolution. Forty-one CSPs were separated and identified from A. catenella DH01 by 2-DE, in which 14 were identified in the protein database using MALDI TOF-TOF MS analysis. This work represents the first attempt to investigate the CSPs of A. catenella using the CyDye DIGE fluor dyeing method that provides a potentially important tool for future comprehensive characterization of CSPs and elucidation of the physiological functions of CSPs in dinoflagellates.Alexandrium catenella, cell surface protein, cyanine dye, protein extraction, two-dimensional gel electrophoresis, MALDI TOF-TOF Citation:Li C, Wang D Z, Dong H P, et al. Proteomics of a toxic dinoflagellate Alexandrium catenella DH01: Detection and identification of cell surface proteins using fluorescent labeling.

The Mechanism of Diarrhetic Shellfish Poisoning Toxin Production in Prorocentrum spp.: Physiological and Molecular Perspectives

“…In addition, RNA and DNA can be extracted simultaneously from the same sample for further analysis. This method has been widely applied in many dinoflagellate species, such as planktonic P. donghaiense [120], PSP toxin-producing A. catenella [121,122,123,124], and YTX-producing Lingulodinium polyedrum [125]. Nevertheless, a few reports have suggested that this method may not produce high-quality results for certain dinoflagellate samples.…”

“…Most proteomic studies in the area of algal toxins have focused on PSP toxin biosynthesis in Alexandrium spp. [118,122,123,124,128,129,130,131]. Certain researchers have compared the 2-DE profiles of 2 strains with considerable variations in the levels of toxins triggered by different environmental conditions or growth phases [124,129,130].…”

Dinoflagellates: Ecological Approaches and Spatial Distributions in Malaysia Waters

“…Dinoflagellates are differentiated from one another by the pattern of thecate or amphiesma which is made up of cellulose plates that protect the cells from the tugor pressure [1,3]. Previous study on the fossils of ancient dinoflagellates revealed six patterns of plate that classified the species into different major groups; gymnodinoid, suessioid, peridinioid, gonyaulacoid, dinophysoid, and prorocentroid [4].…”

Omics in marine biotechnology

“…This topic contains 12 papers produced by research groups from China, Germany, and Korea. To help readers, we have arranged the papers according to their research topics: genomics [1-3]; transcriptomics [4]; proteomics [5][6][7][8][9]; metabolomics [10,11], and nutrigenomics [12].The objective of this topic is to offer a platform for marine biological scientists to present recent advances in the discovery and development of omics in marine biotechnology, providing a large-scale, holistic approach to understanding life in encapsulated "omes". In this special topic, several genomes, including major histocompatibility complex genes [2], phytochrome genes [3] and immune-and antiviral-related genes [4], have been systematically investigated based on functional genomic studies, providing targets for further structural and functional analyses.…”

“…This topic contains 12 papers produced by research groups from China, Germany, and Korea. To help readers, we have arranged the papers according to their research topics: genomics [1-3]; transcriptomics [4]; proteomics [5][6][7][8][9]; metabolomics [10,11], and nutrigenomics [12].…”