优化分离与鉴定蓝斑背肛海兔口腔神经节蛋白质组

Optimized Separation and Identification of Proteome from the Baccal Ganalion of Aplysia(Notarcus leachii cirrosus Stimpson)

采用双向凝胶电泳技术优化分离蓝斑背肛海兔(Notarcus leachii cirrosusStimpson,NLCS)口腔神经节(Buccal Ganglion,BG)蛋白质组,并获得约300个蛋白质斑点.用组合基质辅助激光解吸电离化飞行时间(MALD I-TOF)质谱技术和胶内酶解技术测定BG蛋白质组中的96个蛋白质斑点的肽指纹(Peptide m ass fin-gerprint,PMF)图谱.经数据库检索与比对后,发现96种蛋白质中仅有4种蛋白质可获得较高的匹配率,它们分别是微管蛋白(Tubu lin)、肌动蛋白(Actin)和两个1,5-二磷酸核酮糖-羧化酶/加氧酶(R ibu lose-1,5-b i-sphosphate carboxylase/oxygenase,R ibu lose),均属于神经细胞骨架蛋白质;同时还发现一种交配信号肽前体(Peptide m ating pheromone precursor).利用LOC trees软件和分类法对56种蛋白质进行亚细胞定位与分类.

A separation method of proteome from the buccal ganglion of aplysia （Notarcus leachii cirrosus Stirnpson, 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.