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在镉盐胁迫下用蛋白质组学技术筛选与鉴定海兔亚口腔神经节的差异蛋白质

黄琳1, 陈东仕1, 颜利1, 方财王1, 黄河清1,2,3   

    1. 厦门大学生命科学学院生物化学与生物技术学系,
    2. 海洋与环境科学学院近海海洋科学国家重点实验室,
    3. 化学化工学院化学生物学系, 厦门 361005
  • 收稿日期:2008-06-02 修回日期:1900-01-01 出版日期:2009-02-10 发布日期:2009-02-10
  • 通讯作者: 黄河清

Differential Proteins of Sub-buccal Ganglions Selected and Identified with Proteomic Techniques in Aplysia Under the Stress Condition of Cadmium Chloride

HUANG Lin1,2, CHEN Dong-Shi1, YAN Li1, FANG Cai-Wang1,3, HUANG He-Qing1,2,3*   

    1. Department of Biochemistry and Biotechnology, School of Life Sciences,
    2. State Key Laboratory of Marine Environmental Science, College of Oceangraphy and Environmental Science,
    3. Department of Chemical Biology, College of Chemistry & Chemical Engineering, Xiamen University, Xiamen 361005, China
  • Received:2008-06-02 Revised:1900-01-01 Online:2009-02-10 Published:2009-02-10
  • Contact: HUANG He-Qing

摘要: 以蓝斑背肛海兔(Notarcus leachii cirrosus Stimpson, NLCS)的口腔神经节(Buccal ganglion, BG)为研究对象, 按BG形态对称性, 解剖成亚BG(sub-BG, SBG), 并分为左SBG和右SBG, 简称为LSBG和RSBG. 用双向凝胶电泳(2D-PAGE)技术优化分离LSBG和RSBG全蛋白质, 并采用蛋白质组学和数据库比对技术筛选与鉴定差异蛋白质. 实验结果表明, LSBG和RSBG之间的差异蛋白质主要由活性多肽的前体蛋白或降解后大片段多肽组成, 它们对维持BG的生理功能起着重要的作用. 在急性镉盐(10 μg/mL)胁迫下, NLCS的LSBG和RSBG表达了由镉盐诱导的差异蛋白质, 并采用蛋白质组学技术分别分离、筛选和鉴定, 其主要的差异蛋白质有下调的肌球蛋白、钙结合蛋白、上调的热休克蛋白和硫氧还蛋白. 这些蛋白质可能与BG细胞抗镉毒性有关, 部分差异蛋白质适合于监测镉盐污染且开展毒理学研究的蛋白指示物.

关键词: 海兔, 亚口腔神经节, 蛋白质组学, 镉盐, 蛋白指示物

Abstract: The buccal ganglion(BG) was symmetrically divided into two sub-BG(SBG) in Aplysia(Notarcus leachii cirrosus Stimpson, NLCS) with reference to its ganglion shape, called left SBG(LSBG) and right SBG(RSBG). The whole proteins both LSBG and RSBG were optimally separated via two-dimensional polyacry-lamide gel electrophoresis(2D-PAGE), respectively. The differential proteins were further selected and identified by proteomic and comparison database techniques. The experimental results indicate that these differential proteins between LSBG and RSBG are identified to be the precursor proteins or the large segments of active peptides, which both proteins may play the important role in maintaining the physiological function of BG. Both LSBG and RSBG in NLCS can express the differential proteins induced with the cadmium under the stress of actual cadmium chloride at 10 μg/mL. However, these proteins were effectively separated, selected and identified with proteomic techniques, indicating that these proteins were considered to be the down-regulated proteins such as ropomyosin and Mw=16000 calcium-binding, and the up-regulated proteins such as heat shock protein and thioredoxin. We suggest that these proteins be tightly connected with mithridatism of cadmium in BG cells and , as protein markers in part, be suit for monitoring pollution level of cadmium and developing the research focused on toxicology.

Key words: Aplysia, Sub-baccual ganglion, Proteomics, Cadmium chloride, Protein marker

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