Study of biomolecular complex formation by heteronuclear multidimensional NMR

异核多维核磁共振研究生物分子复合物形成

基本信息

批准号：
09307054
负责人：
KOBAYASHI Yuji
金额：
$ 15.87万
依托单位：
Osaka University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (A)
财政年份：
1997
资助国家：
日本
起止时间：
1997 至 1998
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09307054/
关键词：
complex BMP receptor NMR tertiary structure BMPレセプター構造活性相関複合体形成蛋白質複合体 IGF

项目摘要

Many biomolecules form non-covalent complexes (super molecules) when they are biologically active. In the case of molecules that act in signaling systems, the formation of the non-covalent complex is the initial event that triggers some important biological process. Although many studies have been made on such complexes, little is known about the detailed mechanism of signaling. The purpose of this research project is to explore the structure and dynamics of such protein-protein complexes by multidimensional NMR techniques. Moreover, we also intend to develop physico-chemical methodologies, which are based on sedimentation equilibrium measurements, differential scanning calorimetry and surface plasmon resonance, to investigate the mechanism of complex formation. In particular, we are focusing our attention on the BMP(bone morphogenetic protein) system. We have established a bacterial expression system for BMP and its receptor molecules. The purified recombinant BMP receptor showed high binding affinity, which was analyzed quantitatively by several physico- chemical methods. Site directed mutagenesis studies have revealed that certain amino acid residues of the BMP receptor contribute to complex formation. Using heteronuclear multidimensional NMR methods, the assignments of the BMP receptor are complete and the solution structure of BMP receptor nearly determined. In other words, we have powerful tools and methods to investigate the BMP-BMP receptor complex at high (atomic) resolution.

许多生物分子在生物学活性时形成非共价复合物（超级分子）。对于在信号系统中起作用的分子，非共价复合物的形成是触发一些重要生物学过程的初始事件。尽管已经对这种复合物进行了许多研究，但对信号传导的详细机制知之甚少。该研究项目的目的是通过多维NMR技术探索这种蛋白质蛋白质复合物的结构和动力学。此外，我们还打算开发物理化学方法学，这些方法基于沉积平衡测量，差异扫描量热法和表面等离子体共振，以研究复杂形成的机理。特别是，我们将注意力集中在BMP（骨形态发生蛋白）系统上。我们已经建立了BMP及其受体分子的细菌表达系统。纯化的重组BMP受体显示出较高的结合亲和力，通过几种物理学方法对其进行了定量分析。位点的定向诱变研究表明，BMP受体的某些氨基酸残基有助于复杂的形成。使用异核多维NMR方法，BMP受体的分配是完整的，并且几乎确定了BMP受体的溶液结构。换句话说，我们有强大的工具和方法来研究高（原子）分辨率的BMP-BMP受体复合物。