Expression of cargo receptor ERGIC-53 with defferernt sugar-binding specificities and the effect on carbohydrate structures attached on secretory and membrane proteins.

具有不同糖结合特异性的货物受体 ERGIC-53 的表达以及对附着在分泌蛋白和膜蛋白上的碳水化合物结构的影响。

• 批准号: 11557178
• 负责人: YAMAMOTO Kazuo
• 金额: $ 3.46万
• 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 (B).
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-11557178/
• 关键词: lectin; ERGIC-53; transport; membrane traffic; 糖結合特異性

In secretory pathway newly synthesized secretory and membrane proteins undergo folding and glycosylation during the transport from the ER, through the Golgi. Transport along this pathway sorting of proteins are occurred by cargo receptors. ERGIC-53 is one of the cargo receptors with a leguminous lectin domain on its lumenal side and ER-retrieve signal peptide on its cytoplasmic tail. We constructed the cDNA coding ERGIC-53 substituted of its lectin domain into galactose-binding Bauhinia purpurea lectin (BPA) or sialic acid-binding Maackia amurensis lectin (MAH). Signal peptide on its carboxy terminus was also substituted into those of other trans Golgi localized proteins, TGN38, CI-MPR and furin. Chimeric cargo receptors were expressed in MDCK cells and the effect on the secretory pathway of the cells were analyzed, respectively. Based on the data of western blotting and flow cytometric analyses, chimeric ERGIC-53 with BPA at its lectin domain and TGN38 at cytoplasmic tail transported proteins rich in galactose to the surface of the cells and rich in galactose.

在分泌途径中，新合成的分泌蛋白和膜蛋白在从ER通过高尔基体的运输过程中经历折叠和糖基化。蛋白质的转运沿着这条途径通过货物受体进行分选。ERGIC-53是在其内腔侧具有豆科凝集素结构域并且在其胞质尾区具有ER-检索信号肽的货物受体之一。我们构建了ERGIC-53的cDNA，将其凝集素结构域替换为半乳糖结合的羊蹄甲凝集素（BPA）或唾液酸结合的怀槐凝集素（MAH）。其羧基端的信号肽也被TGN 38、CI-MPR和furin的羧基端信号肽取代。在MDCK细胞中表达嵌合货物受体，并分析其对细胞分泌途径的影响。基于Western印迹和流式细胞术分析的数据，在其凝集素结构域处具有BPA并且在胞质尾处具有TGN 38的嵌合ERGIC-53将富含半乳糖的蛋白质转运至细胞表面并且富含半乳糖。

项目成果

山本一夫: "Chimeric lectin of Bauhinia purpurealectin and Lena curinalistectin recognizes unique carbohydrate structure"J.Biochem.. 127. 129-135 (2000)

Kazuo Yamamoto：“紫荆花凝集素和 Lena curinalistectin 的嵌合凝集素识别独特的碳水化合物结构” J.Biochem.. 127. 129-135 (2000)

山本一夫 他2名: "Cybrorg lectins : Novel leguminous lectins with unique specificities"J,Biochem.. 127(1). 137-142 (2000)

Kazuo Yamamoto 和其他 2 人：“Cybrorg 凝集素：具有独特特异性的新型豆科凝集素”J，Biochem.. 127(1) (2000)。

N.Matsumoto, M.Mitsuki, K.Tajima, W.M.Yokoyama, K.Yamamoto.: "The functional binding site for the c-type lectin-like NK cell receptor Ly49A spans three domains of its MHC class I ligand."J.Exp.Med.. 193. 147-157 (2001)

N.Matsumoto、M.Mitsuki、K.Tajima、W.M.Yokoyama、K.Yamamoto.：“c 型凝集素样 NK 细胞受体 Ly49A 的功能结合位点跨越其 MHC I 类配体的三个结构域。”

山本一夫 他2名: "A chimeric lectin formed from Bauhinia purpurealectin and Lens culinaris lectin recognizes a unique carbohydrate structure"J,Biochem.. 127(1). 129-135 (2000)

Kazuo Yamamoto 和其他 2 人：“由紫荆花凝集素和小扁豆凝集素形成的嵌合凝集素识别独特的碳水化合物结构”J，Biochem.. 127(1) (2000)。

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。”