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Simulation of re-differentiation of plants by transformation using a hormone-like peptide gene

使用类激素肽基因转化模拟植物的再分化

基本信息

批准号：
09460005
负责人：
HIRANO Hisashi
金额：
$ 6.91万
依托单位：
Yokohama City University, Kihara Institute for Biological Research
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1997
资助国家：
日本
起止时间：
1997 至 1999
项目状态：
已结题

项目摘要

A basic 7S globulin isolated from plants is capable of binding to the animal insulin and insulin-like growth factors and shows similarity to the insulin receptor in protein structure, protein kinase activity and subcellular localization , suggesting this protein to have insulin receptor-like functions. A 4-kDa peptide, leginsulin isolated from legumes can bind to the basic 7S globulin. Both leginsulin and the basic 7S globulin are localized in the cell walls and plasma membranes. Leginsulin has stimulatory effect on phosphorylation activity of the basic 7S globulin. Cell proliferation and differentiation of in vitro cultured carrot tissues can be stimulated by adding leginsulin to the medium. The callus tissues of carrot and bird's-foot trefoil transfected the sense leginsulin gene using Agrobacterium transformation system showed rapid growth and formed green spots on the surface. It is concluded that leginsulin is a hormone-like peptide functionally related to animal insulins. The geometries of the disulfide bridges are conserved between leginsulin and insulin. The disulfide bridges of leginsulin are indispensable for stimulatory effect on the phosphorylation. Secondary and tertiary structures of leginsulin are apparently different from those of insulin. However, NMR analysis reveals the similarity in the amino acid residues on the surface of both molecules. The specific binding activity to their binding proteins and the stimulatory effect on protein phosphorylation can be attributed to this similarity. Thus, functionally insulin-like peptides are distributed in not only animals but possibly higher plants.

从植物中分离出一种碱性7S球蛋白，它能与动物胰岛素和胰岛素样生长因子结合，在蛋白质结构、蛋白激酶活性和亚细胞定位等方面与胰岛素受体相似，提示该蛋白具有胰岛素受体样功能。一种从豆类中分离的4-kDa肽，豆类胰岛素可以与碱性7S球蛋白结合。豆类胰岛素和碱性7S球蛋白都定位于细胞壁和质膜。豆类胰岛素对碱性7S球蛋白磷酸化活性有促进作用。在培养液中添加豆类胰岛素可促进胡萝卜组织细胞的增殖和分化。利用农杆菌转化系统将正义豆胰岛素基因转入胡萝卜和百脉根的愈伤组织中，愈伤组织生长迅速，表面形成绿色斑点。结论：豆类胰岛素是一种与动物胰岛素功能相关的类胰蛋白酶肽。二硫键的几何形状在豆类胰岛素和胰岛素之间是保守的。豆胰岛素的二硫键对磷酸化的刺激作用是必不可少的。豆类胰岛素的二级和三级结构与胰岛素的二级和三级结构明显不同。然而，NMR分析揭示了两种分子表面上氨基酸残基的相似性。与其结合蛋白的特异性结合活性和对蛋白磷酸化的刺激作用可归因于这种相似性。因此，功能性胰岛素样肽不仅分布在动物中，而且可能分布在高等植物中。