Construction of cyborg lectins by using lambda foo phage display system, and structural analyses of the lectins complexed with sugars.

利用 lambda foo 噬菌体展示系统构建 cyborg 凝集素，并对与糖复合的凝集素进行结构分析。

• 批准号: 10672038
• 负责人: YAMAMOTO Kazuo
• 金额: $ 1.22万
• 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: 1998
• 资助国家: 日本
• 起止时间: 1998 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10672038/
• 关键词: lectin; phage display system; lambda foo; Bauhinia purpurea lectin; λfooファージ; T7ファージ

Bauhinia purpurea lectin (BPA) is one of β-galactose-binding leguminous lectins. Leguminous lectins contain a long metal-binding loop, the part of which determined of the carbohydrate-binding specificities. In the cDNA coding BPA, random mutations were introduced into a portion of cDNA which corresponds to the carbohydrate-binding loop of this lectin. Mutated lectin library expressed on the surface of lambda foo phages were screened by the panning method. Several phage clones having the affinity for mannose or N-acetylglucosamine were isolated, respectively. These results open new possibilities for making an artificial lectins (named "cyborg lectin") which have distinct and desired carbohydrate-binding specificities.

羊蹄甲凝集素（Bauhinia purpurea lectin，BPA）是一种β-半乳糖结合的豆科植物凝集素。豆科植物凝集素含有一个很长的金属结合环，该环的一部分决定了碳水化合物结合的特异性。在编码BPA的cDNA中，将随机突变引入到对应于该凝集素的碳水化合物结合环的cDNA部分中。用淘选法筛选在λ foo酵母表面表达的突变凝集素文库。分别分离了几个对甘露糖或N-乙酰葡糖胺具有亲和力的噬菌体克隆。这些结果为制备具有独特的和期望的碳水化合物结合特异性的人工凝集素（称为“cyborg凝集素”）打开了新的可能性。

项目成果

S. Iida, K. Yamamoto, T. Irimura.: "Interaction of human macrophage C-type lectin with O-linked N-acetylgalactosamine residues on mucin glycopeptides."J. Biol. Chem.. 274. 10697-10705 (1999)

S. Iida、K. Yamamoto、T. Irimura.：“人巨噬细胞 C 型凝集素与粘蛋白糖肽上 O-连接的 N-乙酰半乳糖胺残基的相互作用。”

S. Kurono, K. Yamamoto, T. Irimura, Y. Nagai.: "Attempts at qualitative and quantitative analysis of noncovalent and specifically bound complexes (protein and sugar chain) by the use of mass spectrometry."Glycoconj. J.. 16. S111 (1999)

S. Kurono、K. Yamamoto、T. Irimura、Y. Nagai.：“尝试使用质谱法对非共价和特异性结合的复合物（蛋白质和糖链）进行定性和定量分析。”Glycoconj。

Y. Iimura, H. Ishikawa, K. Yamamoto and F. Sehnal.: "Hemagglutinating Properties of Apolipophorin III From the Hemolymph of Galleria mellonella Larvae."Arch. of Insect Biochem. and Physiol.. 38. 119-125 (1998)

Y. Iimura、H. Ishikawa、K. Yamamoto 和 F. Sehnal.：“来自大蜡螟幼虫血淋巴的载脂蛋白 III 的血凝特性”。

山本一夫 他4名: "Order and maximum incorporation of N-acetylgalactosamine inte threonine residues of MUC2 core peptide"Biochem. J.. 346(印刷中). (2000)

Kazuo Yamamoto 和其他 4 人：“MUC2 核心肽苏氨酸残基中 N-乙酰半乳糖胺的顺序和最大掺入”Biochem J.. 346（出版中）。

山本一夫: "Analysis of Asparagine-linked oligosaccharides by sequential lectin-affinity chromatography" Methods in Mol.Biol.76. 35-51 (1998)

Kazuo Yamamoto：“通过连续凝集素亲和色谱法分析天冬酰胺连接的寡糖”，Mol.Biol.76 中的方法 (1998)。