Identification of cholera toxin B subunit binding protein and its bioactive analysis

霍乱毒素B亚基结合蛋白的鉴定及其生物活性分析

• 批准号: 18590430
• 负责人: WADA Akihiro
• 金额: $ 2.57万
• 依托单位: Nagasaki University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2006
• 资助国家: 日本
• 起止时间: 2006 至 2007
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18590430/
• 关键词: cholera toxin; CT-B; receptor; immune modulation; GM1

The Cholera toxin (CT)is composed of two subunits, a toxigenic A subunit (CT-A)which activates the adenylyl cyclase system and a pentamaric B subunit (CT-B)which is responsible for CT binding to the cell membrane GM1 gangliosides. The CT is one of the most effective and widely studied mucosal adjuvants. Although the ADP-ribosylating CT-A has been implicated in augmenting immune responses, the receptor-binding CT-B has a greater immunogenicity and may be a repository of adjuvant activity without potential toxicity In general knowledge, the only CT-B receptor is the cell membrane GM1 ganglioside, which expresses in all mammalian cells. The GM1 may be required for the ability of CT-B molecules to alter immunoresponse. However, novel unknown CT-B receptor, which is significant for immune modulation, may account for this signal transduction. In order to elucidate mechanisms of immune modulation by CT-B in vitro, we purified cell surface CT-B binding receptor p32 (32 kDa protein)on SDS-PAGE gel by pull-down method with biotinyl CT-B and avidin-Sepharose. By in-gel digestion of p32 and MALDI-MS fingerprint analysis, p32 was determined as a well-known protein on mammalian cells. The role of CT-B receptor p32 for immune modulation by CT-B will continue to be examined.

霍乱毒素 (CT) 由两个亚基组成，一个是产毒 A 亚基 (CT-A)，可激活腺苷酸环化酶系统；另一个是五元 B 亚基 (CT-B)，负责 CT 与细胞膜 GM1 神经节苷脂的结合。 CT 是最有效且研究最广泛的粘膜佐剂之一。虽然 ADP 核糖基化 CT-A 与增强免疫反应有关，但受体结合的 CT-B 具有更大的免疫原性，并且可能是佐剂活性的储存库，而没有潜在毒性。一般而言，唯一的 CT-B 受体是细胞膜 GM1 神经节苷脂，它在所有哺乳动物细胞中表达。 CT-B 分子改变免疫反应的能力可能需要 GM1。然而，对于免疫调节具有重要意义的新型未知 CT-B 受体可能解释了这种信号转导。为了阐明CT-B在体外的免疫调节机制，我们用生物素CT-B和亲和素-琼脂糖凝胶通过pull-down方法在SDS-PAGE凝胶上纯化了细胞表面CT-B结合受体p32（32 kDa蛋白）。通过 p32 的凝胶内消化和 MALDI-MS 指纹分析，p32 被确定为哺乳动物细胞上众所周知的蛋白质。 CT-B 受体 p32 在 CT-B 免疫调节中的作用将继续得到研究。

项目成果

コレラ毒素Bサブユニットが示す生物活性の解析

霍乱毒素B亚基的生物活性分析

• 发表时间: 2008
• 作者: 和田 昭裕;ら
• 通讯作者: ら

Oncogenesis and the link between inflammation and cancer due to human papillomavirus (HPV) infection, and the development of vaccine control strategies

人乳头瘤病毒 (HPV) 感染引起的肿瘤发生、炎症与癌症之间的联系以及疫苗控制策略的开发

• 发表时间: 2008
• 期刊: Cancer Research Journal (in press)
• 作者: Senba M.;Mori N.;and Wada A.
• 通讯作者: and Wada A.

Helicobacter pylori vacuolating cytotoxin induces activation of the proapoptotic proteirs Bax and Bak, leading to cytochrome c release and cell death, independent of vacuolation

幽门螺杆菌空泡细胞毒素诱导促凋亡蛋白 Bax 和 Bak 的激活，导致细胞色素 c 释放和细胞死亡，与空泡形成无关

• 发表时间: 2006
• 期刊: J Biol Chem 281(16)
• 作者: E.;Yamasaki;A.;Wada;A.;Kumatori;I.;Nakagawa;J.;Funao;M.;Nakayama;J.;Hisatsune;M.;Kimura;J.;Moss;T.;Hirayama
• 通讯作者: Hirayama

Clustering of Helicobacter pylori VacA in lipid rafts, mediated by its receptor, receptor-like protein tyrosine phosphatase beta, is required for intoxication in AZ-521 cells.

幽门螺杆菌 VacA 在脂筏中聚集，由其受体、受体样蛋白酪氨酸磷酸酶 β 介导，是 AZ-521 细胞中毒所必需的。

• 发表时间: 2006
• 期刊: Infect.Immun. 74(12)
• 作者: E.;Yamasaki;A.;Wada;A.;Kumatori;I.;Nakagawa;J.;Funao;M.;Nakayama;J.;Hisatsune;M.;Kimura;J.;Moss;T.;Hirayama;Nakayama M et al.
• 通讯作者: Nakayama M et al.

Oncogenesis and the link between inflammation and cancer due to human papillomavirus(HPV)infection,and the development of vaccine control strategies

人乳头瘤病毒（HPV）感染引起的肿瘤发生、炎症与癌症之间的联系以及疫苗控制策略的开发

• 发表时间: 2008
• 期刊: Cancer Research Joumal (in press)
• 作者: Senba M.;Mori N.;Wada A.
• 通讯作者: Wada A.