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The role of auxin-binding protein during growth and development of fruit

生长素结合蛋白在果实生长发育过程中的作用

基本信息

批准号：
02454046
负责人：
YAMAKI Shohei
金额：
$ 4.86万
依托单位：
Nagoya University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (B)
财政年份：
1990
资助国家：
日本
起止时间：
1990 至 1991
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-02454046/
关键词：
Auxin IAA Auxin-binding protein Protoplast Melon Apple Cymbidium プロトプラスト

项目摘要

Auxin-binding protein in apple seedling was partially purified by 2, 4-D linked-Sephasrose 4B affinity chromatography and 2 times of Sepharose 4B gel filtration. It had a molecular weight of about 400 KDa and was composed of subunit bands of 56 KDa ana 15 KDa by SDS-PAGE, which appeared in proportion to the strength of 6binding activity. The dissociation constant by Scatchard plot was 7.7X1O^<-6> M. This binding protT4n is similar to ABP-I from mung bean seedling. The binding activity to ^<14>C-IAA was completely inhibited by 2, 4-D and NAA, and inhibited to about 50% by PCIB of antiauxin. Auxin-binding protein in melon seedling partially purified by the same procedure showed the equal activity between both assay methods of an ammonium sulfate precipitation and an equilibrating dialysis. The protein had a molecular weight of 155 KDa and a dissociation constant of 4.8X1O^<-6> M, and combined more than one molar IAA to one molar protein. This binding protein from mellon seedling is like ABP-II from mung bean seedling.The function about two kinds of the auxin-binding protein was not investigated in detail through the present experiment. However, we can suggest firmly these auxin binding proteins possess the characteristics as an auxin receptor, as defined by the specificity for substrate, the saturation of reaction to substrate concentration and the reversibility of reaction.

采用2，4-D linked-Sepharose 4 B亲和层析和2次Sepharose 4 B凝胶过滤，部分纯化了苹果幼苗生长素结合蛋白。SDS-聚丙烯酰胺凝胶电泳显示，该蛋白分子量约为400 KDa，由56 KDa和15 KDa两条亚基带组成，亚基带的大小与结合活性的强弱成正比。通过Scatchard图的解离常数为7.7 × 10 - 4 <-6>M。该结合蛋白T4 n与绿豆幼苗的ABP-I相似。2<14>，4-D和NAA完全抑制了该酶与~ 1C-IAA的结合活性，而抗生长素的PCIB则抑制了该酶与~ 1C-IAA的结合活性约50%。用同样的方法部分纯化的甜瓜幼苗生长素结合蛋白在硫酸铵沉淀和平衡透析两种测定方法之间显示出相等的活性。该蛋白质的分子量为155 KDa，解离常数为4.8 × 10 - 4 <-6>M，并且将超过1摩尔的IAA结合到1摩尔的蛋白质中。这两种生长素结合蛋白的功能在本实验中还没有详细的研究。然而，我们可以坚定地认为，这些生长素结合蛋白具有生长素受体的特性，如由底物的特异性，反应对底物浓度的饱和性和反应的可逆性定义的。