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萌发大麦胚的质膜蛋白质组

The plasma membrane proteome of germinating barley embryos.

作者信息

Hynek Radovan, Svensson Birte, Jensen Ole Nørregaard, Barkholt Vibeke, Finnie Christine

机构信息

Enzyme and Protein Chemistry, Department of Systems Biology, Technical University of Denmark, DK-2800 Kgs. Lyngby, Denmark.

出版信息

Proteomics. 2009 Jul;9(14):3787-94. doi: 10.1002/pmic.200800745.

Abstract

Cereal seed germination involves a complex coordination between different seed tissues. Plasma membranes must play crucial roles in coordination and execution of germination; however, very little is known about seed plasma membrane proteomes due to limited tissue amounts combined with amphiphilicity and low abundance of membrane proteins. A fraction enriched in plasma membranes was prepared from embryos dissected from 18 h germinated barley seeds using aqueous two-phase partitioning. Reversed-phase chromatography on C(4) resin performed in micro-spin columns with stepwise elution by 2-propanol was used to reduce soluble protein contamination and enrich for hydrophobic proteins. Sixty-one proteins in 14 SDS-PAGE bands were identified by LC-MS/MS and database searches. The identifications provide new insight into the plasma membrane functions in seed germination.

摘要

谷物种子萌发涉及不同种子组织之间复杂的协调过程。质膜在萌发的协调和执行中必定起着关键作用;然而,由于组织量有限,加之膜蛋白具有两亲性且丰度较低,人们对种子质膜蛋白质组了解甚少。使用双水相分配法从萌发18小时的大麦种子中分离出的胚中制备了富含质膜的组分。采用在微柱上用异丙醇逐步洗脱的方式,在C(4)树脂上进行反相色谱,以减少可溶性蛋白质污染并富集疏水蛋白质。通过LC-MS/MS和数据库搜索鉴定了14条SDS-PAGE条带中的61种蛋白质。这些鉴定结果为种子萌发过程中质膜的功能提供了新的见解。

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