Hynek Radovan, Svensson Birte, Jensen Ole Nørregaard, Barkholt Vibeke, Finnie Christine
Enzyme and Protein Chemistry, Department of Systems Biology, Technical University of Denmark, DK-2800 Kgs. Lyngby, Denmark.
Proteomics. 2009 Jul;9(14):3787-94. doi: 10.1002/pmic.200800745.
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.
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。
