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Optimization of 2D‐PAGE protocols for proteomic analysis of two nonaxenic toxin‐producing freshwater cyanobacteria: Cylindrospermopsis raciborskii and Raphidiopsis sp.
Author(s) -
Plominsky Á.M.,
SotoLiebe K.,
Vásquez M.
Publication year - 2009
Publication title -
letters in applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.698
H-Index - 110
eISSN - 1472-765X
pISSN - 0266-8254
DOI - 10.1111/j.1472-765x.2009.02665.x
Subject(s) - cylindrospermopsis raciborskii , cyanobacteria , toxin , biology , botany , microbiology and biotechnology , ecology , bacteria , genetics
Aims: To optimize a protocol for the extraction and an in‐depth analysis of the soluble protein fraction of two nonaxenic toxin‐producing cyanobacteria Cylindrospermopsis raciborskii (hepatotoxin‐producing), and Raphidiopsis sp. (neurotoxin‐producing), using two‐dimensional polyacrylamide gel electrophoresis (2D‐PAGE). Methods and Results: The soluble protein fractions from strains of C. raciborskii and Raphidiosis sp. with different toxicity phenotypes were analysed by 2D‐PAGE. Specific protocols were optimized specifically for each strain. Between 500 and 700 sharp protein spots were distinguished in a single 4–7 pH range 2D‐PAGE for each cyanobacterium. Comparison of the protein maps of C. raciborskii CS‐505 (a cylindrospermopsin‐producing strain) and Raphidiopsis sp. D9 (saxitoxin‐producing strain) against the nontoxic C. raciborskii strain CS‐509 revealed many unique proteins in each protein map. We confirmed that the resolved proteins were cyanobacterial by identifying three randomly chosen protein spots from a Raphidiopsis sp. strain D9 2D‐PAGE, using high‐performance liquid chromatography (HPLC) tandem mass spectrometry (MS). Conclusions: The 2D‐PAGE conditions presented here provide a robust protocol for proteomic studies in two CYN‐ and STX‐producing model organisms, C. raciborskii and Raphidiopsis sp. Significance and Impact of the Study: We present the first protocols for proteomic analyses of Cylindrospermopsis raciborskii and Raphidiopsis sp.