Structural analysis of BMP and BMP receptor

BMP和BMP受体的结构分析

• 批准号: 11470475
• 负责人: KOBAYASHI Yuji
• 金额: $ 8.26万
• 依托单位: Osaka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B).
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-11470475/
• 关键词: BMP; receptor; structure; NMR; NMR; 立体構造解析

Bone morphogenetic proteins (BMPs) regulate the growth, differentiation and apoptosis of various cells. To elucidate the mechanisms of signal transduction by BMP, we analyzed the structure and interactions of BMP receptor. The extracellular domain of the mouse BMP type-IA receptor was expressed as a fusion protein with thioredoxin in Escherechia coil. Using heteronuclear multidimensional NMR methods, the assignments of the BMP receptor were completed and the solution structure of BMP receptor was determined. In other words, we have obtained powerful tools and methods to investigate the BMP-BMP receptor complex at atomic resolution. Based upon the resulting structure, we have designed and synthesized several mutants of BMP receptor to identify its ligand (i.e. BMP) binding site. CD spectra of the mutant receptors indicated that all the mutants have essentially the native folding structures. Affinities of the mutants to BMP-4 were measured using several physico-chemical methods, surface plasmon resonance sensor (BIACORE), sedimentation equilibrium and NMR.Binding studies have shown that the side chains of Ile52, Glu54, Thr62 and Thr64 at the surface of the three-stranded β-sheet form a high affinity ligand-binding surface together with Phe75 in loop region. These results provide deeper insights for the structural basis of the ligand recognition by BMP receptor.

骨形态发生蛋白（BMPs）调节多种细胞的生长、分化和凋亡。为了阐明BMP的信号转导机制，我们分析了BMP受体的结构和相互作用。小鼠BMP IA型受体的胞外结构域在Escherechia coil中表达为与硫氧还蛋白的融合蛋白。采用异相多维核磁共振方法，对BMP受体进行了归属，确定了BMP受体的溶液结构。换句话说，我们已经获得了强有力的工具和方法来研究BMP-BMP受体复合物在原子分辨率。基于所得到的结构，我们设计并合成了几种BMP受体的突变体，以确定其配体（即BMP）结合位点。突变体受体的CD谱表明，所有的突变体基本上具有天然的折叠结构。结合研究表明，三链β折叠表面的Ile 52、Glu 54、Thr 62和Thr 64侧链与环区的Phe 75形成高亲和力的配体结合表面。这些结果为BMP受体识别配体的结构基础提供了更深入的认识。

项目成果

S.Matusda: "The Role of Calcium-binding Sites in S-modulin Function"Journal of Biological Chemistry. 273. 20223-20227 (1998)

S.Matusda：“钙结合位点在 S-调节蛋白功能中的作用”生物化学杂志。

C.Wang, T.Kai, T.Tomiyama, T.Yoshida, Y.Kobayashi, E.Nishibori, M.Takata, M.Sakata, and H.Shinohara: "A Scandium Carbide Endohedral Metallofullerene : (Sc_2C_2)@C_<84>"ANGEWANDTE CHEMIE. Vol.40. 397-399 (2000)

C.Wang、T.Kai、T.Tomiyama、T.Yoshida、Y.Kobayashi、E.Nishibori、M.Takata、M.Sakata 和 H.Shinohara：“碳化钪内嵌金属富勒烯：(Sc_2C_2)@C_<

T.Ishino: "Interaction of ribosome recycling factor and elongation factor EF-G with E.coil ribosomes studied by the surface plasmon resonance technique"Genes to Cells. 5. 953-963 (2000)

T.Ishino：“通过表面等离子共振技术研究核糖体循环因子和延伸因子 EF-G 与大肠杆菌核糖体的相互作用”《基因到细胞》。

J.Hasegawa, S.Uchiyama, Y.Tanimoto, M.Mizutani, Y.Kobayashi, Y.Sambongi, and Y.Igarashi: "Selected Mutations in a Mesophilic Cytochrome c Confer the Stability of a Thermophilic Counterpart"The Journal of Biological Chemistry. Vol.275. 37824-37828 (2000)

J.Hasekawa、S.Uchiyama、Y.Tanimoto、M.Mizutani、Y.Kobayashi、Y.Sambongi 和 Y.Igarashi：“嗜温细胞色素 c 中的选定突变赋予嗜热对应物的稳定性”生物化学杂志

T.J.Dennis, T.Kai, K.Asato, T.Tomiyama, H.Shinohara, T.Yoshida, Y.Kobayashi, H.Ishiwatari, Y.Miyake, K.Kikuchi, and Y.Achiba: "Isolation and Characterization by ^<13>C NMR Spectroscopy of [84] Fullerene Minor Isomers"Journal of Physical Chermistry A. Vol.

T.J.Dennis、T.Kai、K.Asato、T.Tomiyama、H.Shinohara、T.Yoshida、Y.Kobayashi、H.Ishiwatari、Y.Miyake、K.Kikuchi 和 Y.Achiba：“^ 的分离和表征