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（BMP）调节各种细胞的生长，分化和凋亡。为了阐明BMP信号转导的机制，我们分析了BMP受体的结构和相互作用。小鼠BMP型-IA受体的细胞外结构域表示为融合蛋白，硫氧还蛋白在埃舍雷希亚线圈中。使用异核多维NMR方法，完成了BMP受体的分配，并确定了BMP受体的溶液结构。换句话说，我们获得了在原子分辨率下研究BMP-BMP受体复合物的强大工具和方法。基于所得结构，我们设计并合成了BMP受体的几个突变体，以识别其配体（即BMP）结合位点。突变体的CD光谱认识到所有突变体基本上都具有天然折叠结构。使用几种物理化学方法，表面等离子体共振传感器（BIACORE），沉积等效和NMR。结合研究测量突变体与BMP-4的亲和力。这些结果为BMP受体识别配体识别的结构基础提供了更深入的见解。

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_<

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

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》。