Comparative Proteomic Analysis Reveals Proteins Putatively Involved in Toxin Biosynthesis in the Marine Dinoflagellate Alexandrium catenella

Abstract

Alexandrium is a neurotoxin-producing dinoflagellate genus resulting in paralytic shellfish poisonings around the world. However, little is known about the toxin biosynthesis mechanism in Alexandrium. This study compared protein profiles of A. catenella collected at different toxin biosynthesis stages (non-toxin synthesis, initial toxin synthesis and toxin synthesizing) coupled with the cell cycle, and identified differentially expressed proteins using 2-DE and MALDI-TOF-TOF mass spectrometry. The results showed that toxin biosynthesis of A. catenella occurred within a defined time frame in the G1 phase of the cell cycle. Proteomic analysis indicated that 102 protein spots altered significantly in abundance (P < 0.05), and 53 proteins were identified using database searching. These proteins were involved in a variety of biological processes, i.e., protein modification and biosynthesis, metabolism, cell division, oxidative stress, transport, signal transduction, and translation. Among them, nine proteins with known functions in paralytic shellfish toxin-producing cyanobacteria, i.e., methionine S-adenosyltransferase, chloroplast ferredoxin-NADP+ reductase, S-adenosylhomocysteinase, adenosylhomocysteinase, ornithine carbamoyltransferase, inorganic pyrophosphatase, sulfotransferase (similar to), alcohol dehydrogenase and arginine deiminase, varied significantly at different toxin biosynthesis stages and formed an interaction network, indicating that they might be involved in toxin biosynthesis in A. catenella. This study is the first step in the dissection of the behavior of the A. catenella proteome during different toxin biosynthesis stages and provides new insights into toxin biosynthesis in dinoflagellates.

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].…”

“…[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]. Others have examined differentially expressed proteins, which were observed by comparing the 2-DE profiles of 2 strains with dramatic variations in the levels of toxin [123,128].…”

Comparison of three protein extraction procedures from toxic and non-toxic dinoflagellates for proteomics analysis

“…Therefore, researches on PSTs as well as dinoflagellates became widely executed in the last few decades with different focus. Proteomics is a new research tool in studying dinoflagellates, and has been widely used in recent years (Chan et al 2002(Chan et al , 2004(Chan et al , 2006Wang et al 2013;Huang et al 2015). The combination of two-dimensional electrophoresis (2-DE) and mass spectrometry analysis works effectively in proteomics studies on dinoflagellates (Chan et al 2002) and other organisms.…”

Gene Mining for Environmental Studies and Applications: Examples from Marine Organisms

“…These different transcriptomic analyses of Alexandrium species have recently been enriched by a comparative proteomic approach to directly identify proteins potentially involved in STX biosynthesis in A. catenella (Wang et al, 2013). To conduct these experiments, algae were sampled at different stages of toxin biosynthesis (no toxin synthesis, initial toxin synthesis, and toxin-synthesizing).…”