Structure and function of vertebrate beta-galactoside-binding lecitins

脊椎动物β-半乳糖苷结合卵磷脂的结构和功能

• 批准号: 02680140
• 负责人: KASAI ken-ichi
• 金额: $ 1.34万
• 依托单位: Faculty of Pharmaceutical Sciences, Teikyo University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 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/grant/KAKENHI-PROJECT-02680140/
• 关键词: animal lectin; galactoside-binding lectin; human lectin; chicken lectin; cDNA cloning; recombinant protein; site-directed mutagenesis; ガラクトシド総合レクチン; cDNAリロ-ニング; 組換たんぱく質

Metal-independent beta-galactoside-binding lectins of vertebrates are considered to be involved a variety of important biological regulations such as development of embryo and morphogenesis of tissues in which specific interaction between complex carbohydrates and their counterparts play crucial roles. They are also suggested to be involved in regulation of immune system and metastasis of malignant cells. In the present research, complete amino acid sequence, complete nucleotide sequence of cDNA, expression of recombinant protein in E. coli and the fate during development of embryo of one of chicken isolectin, i. e., 16 kDa lectin, were elucidated. The results made it possible to understand its roles during morphogenesis of various tissues especially that of skin, and also molecular evolution of this lectin family. Effects of substitution of amino acid residues by site-directed mutagenesis of recombinant human 14 kDa lectin showed that a tryptophan and some cystein residues which had been considered to be mostly important were not essential for carbohydrate binding. A cDNA for another human lectin, 29 kDa lectin, was cloned from a library derived from a breast carcinoma. Complete amino acid sequence of a lectin contained in venom of Crotalus was determined in order to know whether it belongs to the metal-independent lectin family or not. The result clearly showed that the snake venom lectin is a member of calcium-dependent lectin family.

脊椎动物的非金属依赖性β-半乳糖苷结合凝集素被认为参与多种重要的生物学调节，如胚胎发育和组织形态发生，其中复杂碳水化合物与其对应物之间的特异性相互作用起着至关重要的作用。它们还被认为参与免疫系统的调节和恶性细胞的转移。本研究对该基因cDNA的全序列、全核苷酸序列、重组蛋白在大肠杆菌中的表达进行了初步研究。大肠杆菌中分离到的一种鸡同种凝集素，即鸡同种凝集素，在胚胎发育过程中的命运。例如，16 kDa凝集素。这些结果使人们有可能了解它在各种组织，特别是皮肤的形态发生过程中的作用，以及该凝集素家族的分子进化。重组人14 kDa凝集素的定点突变的氨基酸残基的取代的影响表明，色氨酸和一些半胱氨酸残基，已被认为是最重要的是不是必不可少的碳水化合物结合。另一种人凝集素，29 kDa凝集素的cDNA克隆自乳腺癌来源的文库。测定了响尾蛇蛇毒中一种凝集素的氨基酸序列，以确定其是否属于非金属依赖性凝集素家族。结果表明蛇毒凝集素属于钙依赖性凝集素家族。

项目成果

Hirabayashi,J.: "Specific isolation by anhydrotrypsin-agarose chromatography of a recombinant protein tagged with an affinity tail arginine at the C-terminus." J.Mol.Recog.3. 204-207 (1990)

Hirabayashi,J.：“通过脱水胰蛋白酶-琼脂糖色谱法对 C 末端带有亲和尾精氨酸标记的重组蛋白进行特异性分离。”

Y.Sakakura: "Structure of chicken 16ーkDa betaーgalactosideーbinding lectin.Complete amino acid sequence,cloning of cDNA,and production of recombinant lectin." J.Biol.Chem.265. 21573-21579 (1990)

Y.Sakakura：“鸡 16-kDa β-半乳糖苷结合凝集素的结构。完整的氨基酸序列、cDNA 的克隆以及重组凝集素的生产。”21573-21579 (1990)。

Sakakura, Y., Hirabayashi, J., Oda, Y, Ohyama, Y., and Kasai, K.: "Structure of chicken 16-kDa beta-galatoside-binding lectin." J. Biol. Chem.265. 21573-21579 (1990)

Sakakura, Y.、Hirabayashi, J.、Oda, Y、Ohyama, Y. 和 Kasai, K.：“鸡 16-kDa β-半乳糖苷结合凝集素的结构”。

Hirabayashi, J., and Kasai, K.: "specific isolation by anhydrotrypsin-agarose chromatography of a recombinant protein tagged with an affinity tail arginine at the C-terminus." J. Mol. Recog.3. 204-207 (1990)

Hirabayashi, J. 和 Kasai, K.：“通过脱水胰蛋白酶-琼脂糖色谱法对 C 末端带有亲和尾精氨酸标签的重组蛋白进行特异性分离。”

Kasai,K.: "Biochemical properties of vertebrate 14K β-galactoside-binding lectins." Advances in Lectin Research. 3. 10-35 (1990)

Kasai, K.：“脊椎动物 14K β-半乳糖苷结合凝集素的生化特性。凝集素研究进展”3. 10-35 (1990)。