Optimized separation and identification of proteome from the baccal ganalion of Aplysia(Notarcus leachii cirrosus Stimpson)

Abstract

A separation method of proteome from the buccal ganglion, of aplysia (Notarcus leachii cirrosus Stimpson, NLCS) with two-dimensional polyacrylamide gel electrophoresis(2D-PAGE) was optimized, gaining about 300 protein spots. Mass spectrogram of peptide mass fingerprint(PMF) of 96 protein spots from the BG were obtained by a combined off-line technique of the matrix-assisted laser desorption ionization mass spectrometry(MALDI-TOF MS) and the emzymolysis in gel. Based on the identification via the databases searches and PMF maps, we found that four of 96 proteins in the gel showed the characteristics with high match scores, which were indicated to be framework proteins tubulin, actin, ribulose-1, 5-bisphosphate carboxylase and oxygenase in neurons cell; meanwhile, a peptide mating pheromone precursor was found. Fifty-six proteins in BG were further classified by a LOC tree software according to the search result of subcellular localization.