Characterization of a protein candidate for the ion channel of influenza C virus and its expression.

丙型流感病毒离子通道候选蛋白的表征及其表达。

基本信息

批准号：
07670337
负责人：
HONGO Seiji
金额：
$ 1.41万
依托单位：
YAMAGATA UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1995
资助国家：
日本
起止时间：
1995 至 1996
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07670337/
关键词：
Influenza C virus M gene CM2 ion channel

项目摘要

The biochemical properties of a second protein (CM2) encoded by RNA segment 6 of influenza C virus were investigated.1.Three forms of CM2 with different electrophoretic mobilities (CM2_0, CM2a and CM2b) were detected in infected cells by immunoprecipitation with antiserum to GST-CM2 fusion protein. 2.A mannose-rich oligosaccharide core is added to unglycosylated CM2_0 (Mr-16,000) to form CM2a (Mr-18,000) and the processing of the carbohydrate chain from high mannose type to complex one converts CM2a into CM2b heterogeneous in electrophoretic mobility (Mr-22,000 to 30,000). 3.Labeling of infected cells with [^3H] palmitic acid showed that CM2 is fatty acylated. The fatty acid bond was sensitive to treatment with hydroxylamine and mercaptoethanol, which indicates a labile thioester-type linkage. 4.The CM2 protein was also found to form disulfide-linked dimers and tetramers on SDS-PAGE under nonreducing conditions. 5.Trypsin treatment of infected cell surface as well as that of microsome vesicles from infected cells followed by immunoprecipitation with antiserum to the GST fusion protein containing the 56 C-terminal amino acid residues of CM2 suggested that this domain is intracellular and exposed on the cytoplasmic side of microsomes. 6.A small amount of CM2 is incorporated into progeny virus particles.These results, altogether, suggest that CM2 is an integral membrane protein with biochemical properties similar to those of influenza A virus M2 and influenza B virus NB proteins.

研究了第二种由甲型流感C病毒RNA第6段编码的第二蛋白（CM2）的生化特性。1。三种形式的具有不同电泳迁移率不同的CM2（CM2_0，CM2A和CM2B）在受感染的细胞中通过与抗抗氧化抗氧化剂至GST-CM2效率蛋白蛋白蛋白蛋白蛋白质在感染的细胞中检测到感染细胞中。 2.将富含甘露糖的寡糖核心添加到未糖基化的CM2_0（MR-16,000）中，以形成CM2A（MR-18,000）（MR-18,000），并将碳水化合物链从高甘露糖类型的加工到复杂的碳水化合物链中，将CM2A转换为CM2A中的CM2B中的CM2B中的CM2B在电动竞争中的CM2B异质（MR-22,000至30,000）。 3.用[^3H]棕榈酸对感染细胞的标签表明CM2是脂肪酰化的。脂肪酸键对用羟胺和墨塔乙醇的治疗敏感，这表明硫酯型不稳定。 4.在非还原条件下，还发现CM2蛋白在SDS-PAGE上形成二硫键连接的二聚体和四聚体。 5.四次治疗感染细胞表面以及来自感染细胞的微型小囊泡，然后用抗血清对含有CM2的56 C末端氨基酸残基的GST融合蛋白进行免疫沉淀，这表明该结构域是内细胞内的，在微质体细胞质的侧面曝光。 6.少量的CM2掺入后代病毒颗粒中。这些结果完全表明CM2是一种积分的膜蛋白，其生化特性与流感A A A A A型病毒M2和流感B病毒NB NB NB NB NB蛋白相似。