NMR PEPTIDE BINDING STUDIES OF PDZ DOMAIN OF CASK

Cask PDZ 结构域的 NMR 肽结合研究

• 批准号: 6279666
• 负责人: AXEL T BRIINGER
• 金额: $ 1.66万
• 依托单位: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-05-01 至 1999-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6279666
• 关键词: biological products; biomedical resource; proteins

CASK is a protein which has been identified to be of importance in cell-cell signaling related to the neurexin neuronal cell surface protein family. The cytoplasmid domain of neurexin proteins interact with a small, 84 residue PDZ domain of CASK. The current project is aimed at understanding, at the molecular level, this binding process. Central to this effort is solving the three dimensional structure of the PDZ domain, which we have done by X-ray crystallography. With the protein structure in hand we now plan to determine the binding conformation of peptides homologous to the presumed binding region of the neurexins by multidimensional heteronuclear NMR. We have made significant progress on this effort. In collaboration with Dr. Chris Turner and Dr. Susan S. Pochapsky we have recorded several 2DH-"N-HSQC spectra on the uncomplexed and peptide bound PDZ domain and have verified that the binding event will significantly perturb amide chemical shifts of a number of key residues. Additionally, we have recorded 3D 1H,"N-filtered NOESY-HSQC and TOCSY-HSQC spectra of the two forms. These spectra will aid significantly in the spectral assignment and the determination of the binding mode of the peptide.

CASK是一种蛋白质，已被鉴定为在 neurexin神经元细胞表面相关的细胞-细胞信号传导 蛋白质家族 neurexin蛋白质的细胞质粒结构域相互作用 具有CASK的小的84个残基PDZ结构域。 本项目属 旨在从分子水平上理解这种结合过程。 这项工作的核心是解决三维结构的 PDZ结构域，我们用X射线晶体学做的。 与 蛋白质结构在手，我们现在计划确定结合 与假定的结合区同源的肽的构象 neurexins的多维异谱NMR。 我们取得了 这一努力取得了重大进展。 与克里斯博士合作 Turner和Susan S. Pochapsky我们已经记录了几个2DH-“N-HSQC 在未复合的和肽结合的PDZ结构域上的光谱， 验证了结合事件将显著干扰酰胺 一些关键残基的化学位移。 此外，我们还有 记录的3D 1H，N-过滤NOESY-HSQC和TOCSY-HSQC光谱 两种形式。 这些光谱将大大有助于光谱分析。 分配和确定肽的结合模式。