STUDY ON THE MODE OF ACTION OF MONOCLONAL NONSPECIFIC SUPPRESSOR FACTOR (MNSF)

单克隆非特异性抑制因子(MNSF)作用方式的研究

• 批准号: 08670367
• 负责人: NAKAMURA Morihiko
• 金额: $ 1.22万
• 依托单位: SHIMANE MEDICAL UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1996
• 资助国家: 日本
• 起止时间: 1996 至 1997
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08670367/
• 关键词: nonspecific; suppressor factor; T cell receptor; サンプレッサー因子; ユビキチン

Hybridoma-derived monoclonal nonspecific suppressor factor (MNSF) was purified with the use of anti-MNSF monoclonal antibody. SDS-polyacryl amide gel electrophoresis of the purified native MNSF was performed under reducing conditions, followed by immunoblot transfer and chemical staining with Amino black. The 70-kDa band was excised, and subjected to amino acid sequence analysis. The N-terminal sequence of 18 amino acids was determined. Degenerated primers corresponding to five amino acids were synthesized and used for RF-PCR.MNSFalpha mRNA was expressed in concanavalin A (Con A) activated (12 hours ) -, but not in unstimulated- T cells. The amino acid sequences deduced from the partial MNSFalpha cDNA showed no homology to any known proteins. However, the sequence showed 35% homology to T cell receptor (TCR) alpha chain in good aggrement with a hypothesis that MNSFalpha might be related to TCRalpha chain. The sequencing of full MNSFalpha cDNA was under way. The elucidation of the mode of action of MNSF with the use of recombinant MNSFalpha should be possible, in the near future.

用抗MNSF单抗纯化杂交瘤来源的非特异性抑制因子(MNSF)。纯化的天然MNSF在还原条件下进行SDS-聚丙烯酰胺凝胶电泳，然后进行免疫印迹转移和氨基黑化学染色。切下70 kDa的条带，进行氨基酸序列分析。测定了18个氨基酸的N末端序列。合成了5个氨基酸对应的简并引物，用于RF-PCR。MNSFpha在刀豆蛋白A(ConA)激活(12h)时表达，但在未刺激的T细胞中不表达。由部分MNSFpha基因推导出的氨基酸序列与任何已知蛋白质都没有同源性。然而，该序列与T细胞受体(TCR)α链有35%的同源性，推测MNSFα可能与TCRα链有关。MNSFpha基因的全序列测定正在进行中。在不久的将来，利用重组的MNSFpha阐明MNSF的作用方式应该是可能的。

项目成果

Morihiko Nakamura: "Ubiquitin-like polypeptide conjugates to acceptor proteins in Concanavalin A-and interferon C-Stimulated T-cells." Biochemical Journal. 330. 683-688 (1998)

Morihiko Nakamura：“泛素样多肽与伴刀豆球蛋白 A 和干扰素 C 刺激的 T 细胞中的受体蛋白结合。”

Morihiko Nakamura: "Ubiquitin-like polypeptide inhibits the IgE response of lipopolySacckaride-activated B cells" International Immunology. 8. 1659-1665 (1996)

Morihiko Nakamura：“泛素样多肽抑制脂多糖激活的 B 细胞的 IgE 反应”国际免疫学。

Morihiko Nakamura: "Ubiquitin-like polypeptide conjugates to acceptor proteins in concanavalin A-and interferon r-stimulated T-cells" Biochemical Journal. 330. 683-688 (1998)

Morihiko Nakamura：“泛素样多肽与伴刀豆球蛋白 A 和干扰素 r 刺激的 T 细胞中的受体蛋白缀合”《生化杂志》。

Yuko Nariai, Morihiko Nakamura, Tomoyoshi Kondoh and Yoshinori Tanigawa: "Production and characterization of a monoclonal antibody specific for ubiquitin-like polypeptide." Journal of Immunoassay. (in pres). (1998)

Yuko Nariai、Morihiko Nakamura、Tomoyoshi Kondoh 和 Yoshinori Tanikawa：“泛素样多肽特异性单克隆抗体的生产和表征。”

Morihiko Nakamura: "Ubiquitin-like polypeptide inhibits the IgE response of lipopolysaccharide-activated B cells" International Immunology. 8. 1659-1665 (1996)

Morihiko Nakamura：“泛素样多肽抑制脂多糖激活的 B 细胞的 IgE 反应”国际免疫学。