华东师范大学学报(自然科学版) ›› 2005, Vol. 2005 ›› Issue (2): 91-97.

• 生命科学 环境科学 • 上一篇    下一篇

液泡前体蛋白质的双向电泳分离和质谱分析

莫蓓莘1,姜里文2   

  1. 1.深圳大学生命科学学院, 深圳518060;2.香港中文大学生物系,新界 香港
  • 收稿日期:2004-04-18 修回日期:2004-09-15 出版日期:2005-05-25 发布日期:2005-05-25
  • 通讯作者: 莫蓓莘

Identification of Proteins from Tobacco Prevacuolar Compartment by Two-Dimensional Electrophoresis and Peptide Mass Fingerprinting(Chinese)

MO Bei-xin1,JIANG Li-wen2   

  1. 1.Life Science Institute, Shenzhen University, Shenzhen518060,China;2.Department of Biology, The Chinese University of Hong Kong, Hong Kong,China
  • Received:2004-04-18 Revised:2004-09-15 Online:2005-05-25 Published:2005-05-25
  • Contact: MO Bei-xin

摘要： 该文通过改进双向电泳方法克服了膜蛋白由于有疏水性而很难进入双向电泳第一相——等电聚焦电泳凝胶，以及由于从蔗糖密度梯度离心中分离得到的液泡前体富含干扰等电聚焦电泳效果的蔗糖等两大难点，成功地分离了通过蔗糖密度梯度离心分离得到的液泡前体蛋白，并使膜蛋白进入了双向电泳凝胶.双向电泳后分离到约200余个蛋白质点，用基质辅助激光解吸/电离飞行时间串联质谱(MS/MS)进行肽质量指纹谱分析,并通过数据库检索进行蛋白质鉴定与功能预测,分析的23个蛋白中有13个在数据库中没有找到相对应的吻合蛋白，查询到的10个吻合蛋白中有6个功能未知.

关键词: 双向电泳, 肽质量指纹谱, 液泡前体, 膜蛋白, 烟草, 双向电泳, 肽质量指纹谱, 液泡前体, 膜蛋白, 烟草

Abstract: It is difficult for membrane proteins to get into 2-D gel because of their hydrophobic properties and the contaminations of carbohydrates in protein samples also affect the results of 2-D gel. The modified 2-D electrophoresis technique described in this paper overcomes these two difficulties, and have successfully separated proteins from sucrose gradient ultracentrifugation and got membrane proteins into 2-D gel. Proteomic analysis was then performed on proteins from prevacuolar compartment. Around 200 protein spots were separated on the 2-D gel, 23 protein spots were cut out form the gel and subjected to MS/MS analysis. After database searching, 13 proteins did not hit matching proteins, with the 10 proteins that hit matching proteins, 4 of them have known protein function, 6 of which turned out to be unknown or unnamed proteins.

Key words: peptide mass finger print, prevacuolar compartment, membrane protein, tobacco, 2-D electrophoresis, peptide mass finger print, prevacuolar compartment, membrane protein, tobacco

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