Molecular Cloning of E-Selectin-Glycoprotein Counter Ligand, gp150.

E-选择素-糖蛋白反配体，gp150 的分子克隆。

• 批准号: 11670147
• 负责人: NAKAMURA Mitsuru
• 金额: $ 2.18万
• 依托单位: JICHI MEDICAL SCHOOL
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 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/grant/KAKENHI-PROJECT-11670147/
• 关键词: E-Selectin; Sialyl-LeX; Counter Ligand; Cell Adhesion; Sialomucin Family; O-Linked Glycoprotein; Sialyl-LeX; Sialomucin Family; O-linked glycoprotein

In human pre-B leukemia cells sialyl-LeX (sLeX) antigen is not expressed on every protein but expressed on a specific core glycoprotein with molecular size of 150kDa (gp150). In addition, cell surface sLeX antigen expression level is controlled by core 2 N-acetylglucosaminyltransferase (C2GnT) gene. In addition, we have revealed the followings.1. Not only in pre-B cell differentiation but also in tonsillar B cell activation, C2GnT regulates cell surface sLeX expression level.2. Anti-sLeX mAb reactivity of gp150 was disappeared after O-sialoglycoprotein-endopeptidase (from Pasteurella haemolytica) treatment. This suggests gp150 must be a sialomucin type glycoprotein molecule that has abundant O-linked sugar chains with sialic acid on their termini.3. A tryptic peptide was purified and analyzed from pre-B NALL-1 cell line membranous fraction using anti-sLeX mAb-conjugated protein-G.The peptide was determined as a mucin-type glycoprotein. It was confirmed by immunostaining methods.4. The cDNA of gp150 was obtained and transfected in the antisense orientation into a sLeX-highly expressed pre-B cell line. Sialyl-LeX-reactivity in the transfected cell line was significantly downregulated and E-selectin-dependent cell adhesion capability was remarkably reduced.

在人前B白血病细胞中，sialyl-LeX（sLeX）抗原并不表达在每种蛋白上，而是表达在分子大小为150 kDa的特异性核心糖蛋白（gp 150）上。此外，细胞表面sLeX抗原表达水平受核心2 N-乙酰葡糖胺转移酶（C2 GnT）基因控制。此外，我们还揭示了以下内容。1. C2 GnT不仅在前B细胞分化过程中，而且在扁桃体B细胞活化过程中调节细胞表面sLeX的表达水平.在O-唾液酸糖蛋白-内肽酶（来自溶血性巴斯德氏菌）处理后，gp 150的抗sLeX mAb反应性消失。这表明gp 150是一种唾液粘蛋白型糖蛋白分子，它含有大量的O-糖链，糖链末端含有唾液酸.胰蛋白酶肽纯化和分析前B NALL-1细胞系膜级分使用抗sLeX单克隆抗体结合蛋白G。该肽被确定为粘蛋白型糖蛋白。免疫组化方法证实.获得gp 150的cDNA并以反义方向转染入sLeX高表达的前B细胞系。转染细胞系中的唾液酸LeX反应性显著下调，E-选择素依赖的细胞粘附能力显著降低。

项目成果

Yang,W.,Asakura,S.,Sakai,T.,Nakamura,M., et al.: "Two-step Spreading Mode of Human Glioma Cells on Fibrin Monomer via a β1 Integrin."Thromb.Res.. 93. 279-290 (1999)

Yang, W.、Asakura, S.、Sakai, T.、Nakamura, M. 等人：“人胶质瘤细胞通过 β1 整合素在纤维蛋白单体上的两步扩散模式。” 93。 279-290 (1999)

中村充: "接着分子シアリルLewis-Xの発現調節"臨床免疫. 31(5). 616-622 (1999)

Mitsuru Nakamura：“粘附分子唾液酸路易斯-X 的表达调节”临床免疫学 31(5) (1999)。

Muroi K, Tarumoto T, Akioka T, Kirito K, Nagai T, Izumi T, Nakamura M, Hatake K, Hakomori S, Miura Y, and Ozawa K.: "Sialyl-Tn- and Neuron Specific Enolase-Positive Minimally Differentiated Erythroleukemia."Internal Medicine. 39(10). 843-846 (2000)

Muroi K、Tarumoto T、Akioka T、Kirito K、Nagai T、Izumi T、Nakamura M、Hatake K、Hakomori S、Miura Y 和 Ozawa K.：“唾液酸-Tn-和神经元特异性烯醇化酶阳性微分化红白血病。

Nakamura,M., et al.: "UDP-GlcNAc:Galβl→3GalNAc βl→6N-acetylglucosaminyltrans-ferase Holds a Key Role on the Control of CD 15s Expression in Human pre-B Lymphoid Cell Lines."Glycobiology. 9(1). 1-12 (1999)

Nakamura, M., 等人：“UDP-GlcNAc:Galβ1→3GalNAc β1→6N-乙酰葡糖胺基转移酶在人前 B 淋巴细胞系中 CD 15 表达的控制中发挥着关键作用。”糖生物学 9(1)。 ）1-12（1999）。

Furukawa,Y.,Iwase,S.,Kikuchi,J.,Nakamura,M., et al.: "Transcriptional Repression of the E2F-1 Gene by Interferon-a Is Mediated through Induction of E2F-4/pRB and E2F-4/pl30 Complexes."Oncogene. 18. 2003-2014 (1999)

Furukawa,Y.,Iwase,S.,Kikuchi,J.,Nakamura,M., et al.：“干扰素-a 对 E2F-1 基因的转录抑制是通过 E2F-4/pRB 和 E2F- 的诱导介导的”