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Novel mechanism of action of monoclonal nonspecific suppressor factor (MNSF)

单克隆非特异性抑制因子（MNSF）的新作用机制

基本信息

批准号：
13680715
负责人：
NAKAMURA Morihiko
金额：
$ 2.11万
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：
2001
资助国家：
日本
起止时间：
2001 至 2002
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13680715/
关键词：
Ubiquitin Suppressor Signal transduction T細胞受容体

项目摘要

Monoclonal nonspecific suppressor factor (MNSF), a lymphokine produced by murine T cell hybridoma, possesses pleiotrophic antigen-nonspecific suppressive function. We have shown that 70-kDa MNSF comprises an 8-kDa ubiquitin-like polypeptide (Ubi-L) and 62-kDa TCRα-like molecule. Ubi-L binds specifically to its 82-kDa receptor protein on target cells. In the current study, We have further characterized the biochemical nature of the TCRa-like molecule. The 62-kDa protein was separated into two species of 46-kDa and 16-kDa on reverse-phase HPLC. Anti-TCRα monoclonal antibody recognized the 46-kDa, but not 16-kDa protein. Anti-TCRβ monoclonal antibody failed to recognize these proteins. Ubi-L conjugated to the 46-kDa protein, whereas Ubi-L lacking its C-termiai Gly-Gly did not. Although Ubi-L was labile both to heating at 56℃ and to acidification to pH 4, Ubi-L46-kDa protein complex was unaffected by these treatments. In addition, the 46-kDa protein elongated the Ubi-L-induced protein tyrosine phosphorylations in concanavalin A-activated murine T helper type 2 clone, D10 cells. One of the four tryptic peptide sequences derived from the 46-kDa protein was in alignment with a related sequence found in the Ja regjon of the TCRa, including the highly conserved motif F-G-X-G-T-X-L.

单克隆非特异性抑制因子（Monoclonal nonspecific suppressor factor， MNSF）是一种由小鼠T细胞杂交瘤产生的淋巴因子，具有多养性抗原非特异性抑制功能。我们已经证明，70 kda的MNSF包含一个8 kda的泛素样多肽（Ubi-L）和62 kda的tcr α样分子。Ubi-L与靶细胞上的82-kDa受体蛋白特异性结合。在目前的研究中，我们进一步表征了tra样分子的生化性质。反相高效液相色谱法将62-kDa蛋白分离为46-kDa和16-kDa两种。抗tcrα单克隆抗体识别46-kDa蛋白，但不识别16-kDa蛋白。抗tcrβ单克隆抗体不能识别这些蛋白。Ubi-L与46-kDa蛋白结合，而Ubi-L缺乏其c -末端，而Gly-Gly则没有。虽然Ubi-L对56℃加热和酸化至pH 4都不稳定，但Ubi-L46-kDa蛋白复合物不受这些处理的影响。此外，46-kDa蛋白在concanavin a激活的小鼠2型T辅助克隆D10细胞中延长了ubi - l诱导的蛋白酪氨酸磷酸化。从46-kDa蛋白中获得的四个色氨酸肽序列中有一个与TCRa Ja区中发现的相关序列一致，包括高度保守的基序F-G-X-G-T-X-L。