Domain structure, expressional regulation and autoimmunity of HSP90

HSP90的结构域结构、表达调控和自身免疫

• 批准号: 10671746
• 负责人: NEMOTO Takayuki
• 金额: $ 2.24万
• 依托单位: Nagasaki University (1999)Iwate Medical University (1998)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10671746/
• 关键词: stress protein; molecular chaperone; HSP90; domain; systemic lupus erythematosus (SLE); oligomer; dimer; 自己抗体; ドメイン構造; 骨芽細胞; アイメフォーム

By use of isoform-specific monoclonal antibodies, the expression of the two HSP90 isoforms was investigated on various rat tissues and osteoclasts and osteoblasts that were induced in alveolar bones of the experimental movement of rat molar teeth. As a result, the predominant and constitutive distribution of HSP90β and stress-induced expression of HSP90α in most murine tissues were demonstrated. We also investigated the domain structure of HtpG, an Esherichia coli homologue of mammalian HSP90 and its domain-domain interactions. HtpG had three major cleavage sites, Arg7-Gly8, Arg336-Glu337 and Lys552-Leu553, susceptible to trypsin. Thus, HtpG consists of three domains: Domain A, Metl-Arg336; Domain B, Glu337-Lys 552; Domain C, Leu553-Ser624. Three kinds of interactions work between the domains of a HtpG dimer: Domain B interacted both with Domain A and Domain C, and moreover, Domain B had a homodimeric interaction. The interaction between Domain B and Domain C was responsible for the dimer conformation of HtpG. Furthermore, Domain B was functionally and structurally divided into two parts: the N-terminal two-third (Glu337-Phe480) that interacted with Domain A and the C-terminal one-third (G1n481-Lys552) that interacted with Domain C. This study first demonstrated the domain structure of HtpG and the interactions between the domains. These characteristics seem to be common among HSP90-family member proteins.

通过使用异构体特异性单克隆抗体，两种HSP 90异构体的表达进行了研究，在各种大鼠组织和破骨细胞和成骨细胞，诱导大鼠磨牙实验移动的牙槽骨。结果表明，HSP90 β在大部分小鼠组织中的优势分布和组成性分布以及应激诱导的HSP90 α表达。我们还研究了哺乳动物HSP90的大肠杆菌同源物HtpG的结构域结构及其结构域之间的相互作用。HtpG有三个主要的酶切位点，Arg7-Gly8，Arg336-Glu337和Lys552-Leu553，对胰蛋白酶敏感。因此，HtpG由三个结构域组成：结构域A，Metl-Arg 336;结构域B，Glu 337-Lys 552;结构域C，Leu553-Ser 624。结构域之间存在三种相互作用：结构域B与结构域A和结构域C均相互作用，结构域B具有同源二聚体相互作用。结构域B和结构域C之间的相互作用是导致HtpG二聚体构象的原因。结构域B在功能和结构上分为两部分：N端三分之二（Glu337-Phe480）与结构域A相互作用，C端三分之一（G1n481-Lys552）与结构域C相互作用。本研究首次揭示了HtpG的结构域结构以及结构域之间的相互作用。这些特征似乎在HSP90家族成员蛋白中是共同的。

项目成果

Maruya M.: "Monomer arrangement in HSP90 dimer as determined by decoration with N- and C-terminal specific antibodies"Journal of Molecular Biology. 285. 903-907 (1999)

Maruya M.：“通过 N 端和 C 端特异性抗体的修饰确定 HSP90 二聚体中的单体排列”分子生物学杂志。