BIOLOGICAL MEANING OF MAJOR CYTOKINININ COCONUT MILK.

主要细胞分裂素椰奶的生物学意义。

• 批准号: 09680564
• 负责人: KOBAYASHI Hisayoshi
• 金额: $ 2.43万
• 依托单位: THE UNIVERSITY OF TOKYO
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09680564/
• 关键词: CYTOKININ; COCONUT MILK; GAZER; STRUCTURE; ZEATIN; CSBP; G3R2-ZR; 糖鎖

A cytokinin isolated from the fluid endosperm of Cocos mucifera L.(coconut milk), accounting for more than 20% of the total cytokinin activity, was structurally analyzed by NMR techniques, mass spectrometry, and sugar analysis by high performance liquid chromatography (HPLC). The planar structure of the cytokinin was deduced from its NMR and mass spectrometric data. The structure of the sugar moiety, including its absolute structure, was determined by HPLC analysis of alditol acetates and aldononitrile acetates derived from the cytokinin. The configuration of the sugar-sugar bonds was determined by NMR, and the structure was finally identified as 14-O-{3-O-[beta-D-galactopyranosyl-(l*2)-alpha-D-galactopyranosyl-(1*3)-alpha-L-arabinoffuranosyl]-4-O-(alpha-L-arabinofuranosyl)-beta-D-galactopyranosyl}-trans-zeatin riboside (GAZER).Cytokinin-specific binding protein (CSBP) was purified from Mung Bean (Vigna radiata) by the use of N-phenyl-N' -(4-pyridyl)urea (4PU)-Sepharose 4B, an affinity gel liganded with 4PU.We determined partial amino acid sequences of CSBP and isolated its cDNA by reverse-transcription (RT) PCR.The cDNA encoded a protein with a calculated molecular mass of 17 kDa. A data based homology search revealed that CSBP in a novel member of a major pollen allergen/pathogenesis-related protein family.

从椰子汁胚乳中分离到一种细胞分裂素，经核磁共振、质谱分析和高效液相色谱糖分析等手段对其进行了结构分析，其中细胞分裂素的活性占总活性的20%以上。根据核磁共振和质谱学数据推测了该细胞分裂素的平面结构。通过对细胞分裂素衍生的阿迪醇乙酸酯和乙醛缩腈的高效液相色谱分析，确定了糖基的结构，包括其绝对结构。用N-苯基-N‘-(4-吡啶基)尿素(4PU)-Sepharose4B从绿豆(Vigna Radiata)中分离纯化了细胞激动素特异性结合蛋白(14-O-{3-O-[beta-D-galactopyranosyl-(l*2)-alpha-D-galactopyranosyl-(1*3)-alpha-L-arabinoffuranosyl]-4-O-(alpha-L-arabinofuranosyl)-beta-D-galactopyranosyl}-trans-zeatin-SPBR)。用4PU连接的亲和凝胶测定了CSBP的部分氨基酸序列，并用逆转录-聚合酶链式反应(RT-PCR)分离了CSBP的cDNA，其编码的蛋白质的相对分子质量为17 kDa。基于数据的同源性搜索显示，CSBP是一个主要的花粉变应原/病程相关蛋白家族的新成员。

项目成果

Hisayoshi Kobayashi: "Strctural identification of a major cytokinin in coconut milk as 14-O-{3-O-{β-D-galactopyranosvl-(1→2)-α-D-galactopyranosyl-(1→3)-α-L-arabinofuranosyl]-4-O-(α-L-arabinofuranosyl)-β-D-galactopyranosyl}-trans-zeatin riboside." Chem.Pha

Hisayoshi Kobayashi：“椰奶中主要细胞分裂素的结构鉴定为 14-O-{3-O-{β-D-吡喃半乳糖基-(1→2)-α-D-吡喃半乳糖基-(1→3)-α-化学法

Hisayoshi Kobayashi: "Strctural identification of a major cytokinin in coconut milk as 14-O-{3-O-〔β-D-galac to pyranosyl-(1→2)-α-D-galactopyranosyl-(1→3)-α-L-arabinofuranosyl〕-4-O-(α-L-arabinofuranosyl)-β-D-galactopyranosyl}-trans-zeatin riboside." Chem.P

Hisayoshi Kobayashi：“椰奶中主要细胞分裂素的结构鉴定为 14-O-{3-O-[β-D-半乳糖至吡喃糖基-(1→2)-α-D-吡喃半乳糖基-(1→3)- α-L-阿拉伯呋喃糖基]-4-O-(α-L-阿拉伯呋喃糖基)-β-D-吡喃半乳糖基}-反式玉米素核苷。" Chem.P

Hisayoshi Kobayashi, Naoko Morisaki, Yoshitaka Tago, Yuichi Hashimoto, Sigeo Iwai, Emiko Kawachi, Ryuji Nagata and Koichi Shudo: "Strctural identification of a major cytokinin in coconut milk as 14-O- {3-O- [beta-D-galactopyranosyl- (1*2) -alpha-D-galacto

Hisayoshi Kobayashi、Naoko Morisaki、Yoshitaka Tago、Yuichi Hashimoto、Sigeo Iwai、Emiko Kawachi、R​​yuji Nagata 和 Koichi Shudo：“椰奶中主要细胞分裂素 14-O- {3-O- [β-D-吡喃半乳糖基”的结构鉴定