高等学校化学学报

高等学校化学学报

• 研究论文 • 上一篇    下一篇

优化分离与鉴定蓝斑背肛海兔口腔神经节蛋白质组

陈东仕1; 黄河清1,2,3; 吴韩志1,3; 蔡宗苇3,4   

  1. 1. 厦门大学生命科学学院细胞生物学与肿瘤细胞工程教育部重点实验室,
    2. 近海海洋科学国家重点实验室,
    3. 固体表面物理化学国家重点实验室, 厦门 361005;
    4. 香港浸会大学化学系, 香港
  • 收稿日期:2005-06-13 修回日期:1900-01-01 出版日期:2006-07-10 发布日期:2006-07-10
  • 通讯作者: 黄河清

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

CHEN Dong-Shi1; HUANG He-Qing1,2,3*; WU Han-Zhi1,3; CAI Zong-Wei3,4   

  1. 1. The Key Laboratory of MOE for Cell Biology and Tumor Cell Engineering,
    School of Life Science,
    2. State Key Laboratory of Marine Environmental Science,
    3. State Key Laboratory for Physical Chemistry of the Solid Surface, Xiamen
    University, Xiamen 361005, China;
    4. Department of Chemistry, Hong Kong Baptist University, Hong Kong, China
  • Received:2005-06-13 Revised:1900-01-01 Online:2006-07-10 Published:2006-07-10
  • Contact: HUANG He-Qing

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被引次数

19

摘要： 采用双向凝胶电泳技术优化分离蓝斑背肛海兔(Notarcus leachii cirrosus Stimpson, 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(MALDI-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 mass fingerprint, 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(Tubulin)、 肌动蛋白(Actin)和两个1,5-二磷酸核酮糖-羧化酶/加氧酶(Ribulose-1,5-bisphosphate carboxylase/oxygenase, Ribulose), 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 mating pheromone precursor). 利用LOCtrees软件和分类法对56种蛋白质进行亚细胞定位与分类.

关键词: 海兔, 口腔神经节, 蛋白质组, 双向凝胶电泳, 质谱

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.

Key words: Aplysia, Buccal ganglion, Proteome, 2D-PAGE, Mass spectrometry

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