Cloning and Subcellular Localization of GalNAc Transferase

GalNAc 转移酶的克隆和亚细胞定位

• 批准号: 08044299
• 负责人: NISHIMORI Isao
• 金额: $ 0.64万
• 依托单位: Kochi Medical School
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for international Scientific Research
• 财政年份: 1996
• 资助国家: 日本
• 起止时间: 1996 至 无数据
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08044299/
• 关键词: Glycosyltransferase; Immunohistochemistry; N-acetylgalactosamine; cDNA; Pancreas cancer; Monoclonal antibody; Gastric cancer; Colon cancer; 免疫組織科学; 膵臓

1. We have obtained a cDNA clone of the human homologue of the UDP-N-acctylgalactosamine : polypeptide N-acetylgalactosaminyltransferase (GalNAc transferase). AcDNA sequence for human GalNAc transferase showed high homology with its bovine homelogue and was identical to human isozyme Tlthat was recently reported.2. Twenty-three residues peptide was synthesized based on the obtained cDNA sequence and a mouse monoclonal antibody to the peptide (GNT-1) was successfully produced.3. GNT-1 reacted with ductal epithelial cells of the exocrine organs including pancreas, salivary gland, bronchial gland, and mammary gland. Oherewise, GNT-1 had no reactivity with mucus cells of salivary gland and stomach. These results suggest that ductal epithelial cells and mucus cells have different types of GalNAc transferase. It was noteworthy that parietal cells of stomach and a large number of cells in Langerhans islands of pancreas showed nice reactivity with GNT-1, suggesting these cells produce O-glycosylated protein except for mucin.4. Western immunoblotting with GNT-1 showed that the expression of GalNAc transferase T1 varies not only among organ types and cell type, but also according to cell differentiation.5.Based on the results deceribed above, we plan to study the expression of GalNAc transferase T1 in various kinds of gastrointstinal disease and produce a monoclonal antibody to GalNAc transferase T2

1.我们获得了UDP-N-乙酰半乳糖胺的人类同源物的cDNA克隆：多肽N-乙酰半乳糖胺基转移酶（GalNAc转移酶）。人GalNAc转移酶的AcDNA序列与其牛同源酶具有较高的同源性，并且与最近报道的人同工酶T1相同。 2．根据获得的cDNA序列合成了23个残基的肽，并成功制备了该肽的小鼠单克隆抗体（GNT-1）。 3． GNT-1与外分泌器官的导管上皮细胞发生反应，包括胰腺、唾液腺、支气管和乳腺。另外，GNT-1与唾液腺和胃的粘液细胞没有反应性。这些结果表明导管上皮细胞和粘液细胞具有不同类型的 GalNAc 转移酶。值得注意的是，胃壁细胞和胰腺朗格汉斯岛的大量细胞与GNT-1表现出良好的反应性，表明这些细胞产生除粘蛋白外的O-糖基化蛋白。4。 GNT-1的Western免疫印迹显示GalNAc转移酶T1的表达不仅在器官类型和细胞类型之间存在差异，而且还根据细胞分化而变化。 5.基于上述结果，我们计划研究GalNAc转移酶T1的表达多种胃肠道疾病并产生针对 GalNAc 转移酶 T2 的单克隆抗体

项目成果

Nishimori I: "A monoclonal antibody against human UDP-GalNAc : polypeptide N-acetylgalactosaminyltransferase and its reactivity with pancreatic and colon tumor cells." Gastroenterology. 110. A568- (1996)

Nishimori I：“针对人 UDP-GalNAc 的单克隆抗体：多肽 N-乙酰半乳糖氨基转移酶及其与胰腺和结肠肿瘤细胞的反应性。”

Nishimori,Isao: "A monoclonal antibody against human UDP-GalNAc : polypeptide N-acetyl-galactosaminyltransferase and its reactivity with pancreatic and colon tumor cells." Gastroenterology. 110. A568- (1996)

Nishimori,Isao：“针对人 UDP-GalNAc 的单克隆抗体：多肽 N-乙酰基-半乳糖胺基转移酶及其与胰腺和结肠肿瘤细胞的反应性。”

Nishimori I,Kohsaki T,Morita M,Akizawa Y,Sano S,Onishi S,Hollingsworth MA.: "A monoclonal antibody against human UDP-GalNAc : polypeptide N-acetylgalactosaminyltransferase and its reactivity with pancreatic and colon tumor cells." Gastroenterology. 110. A

Nishimori I、Kohsaki T、Morita M、Akizawa Y、Sano S、Onishi S、Hollingsworth MA.：“针对人 UDP-GalNAc 的单克隆抗体：多肽 N-乙酰半乳糖氨基转移酶及其与胰腺和结肠肿瘤细胞的反应性。”