NMR CHARACTERIZATION OF GALECTIN 3 LIGAND INTERACTIONS

这个子项目是利用资源的许多研究子项目之一。 由NIH/NCRR资助的中心拨款提供。对子项目的主要支持 子项目的首席调查员可能是由其他来源提供的， 包括美国国立卫生研究院的其他来源。为子项目列出的总成本可能 表示该子项目使用的中心基础设施的估计数量， 不是由NCRR赠款提供给次级项目或次级项目工作人员的直接资金。 Galectin-3是一种半乳糖结合的哺乳动物蛋白，被认为参与了多种过程，包括细胞内转运、细胞-细胞和细胞-基质相互作用，这些过程调节免疫细胞在慢性炎症和恶性细胞转移中的作用。因此，从结构上表征Galectin-3的配体相互作用和结合配体几何构型，作为设计可能的抗炎和抗癌药物的基础，具有重要意义。标准的核磁共振方法，例如那些根据转移的NOE数据给出质子间距离限制的方法，产生了一些有用的结果，但当涉及碳水化合物配体时，限制的数量可能很少，而且当使用设计的高结合常数和低交换速率的抑制剂时，许多标准方法都失败了。这个合作项目使用新的核磁共振方法提供了额外的定向限制，该方法测量了Galectin-3的过氚和稀土标记的碳水化合物识别结构域上的残余偶极耦合(RDC)、顺磁驰豫增强(PRES)和伪接触位移(PCS)。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Galectin-3 is a galactose-binding mammalian protein with suggested involvement in a variety of processes, including introcellular trafficking, cell-cell and cell-matrix interactions, which modulate immune cells in chronic inflammation and metastasis of malignant cells. Therefore, there is substantial interest in structurally characterizing ligand interactions and bound-ligand geometries of galectin-3 as a basis for the design of possible anti-inflammatory and anti-cancer drugs. Standard NMR methodologies, such as those giving inter-proton distance constraints from transferred NOE data, yield some useful results, but numbers of constraints can be small when carbohydrate ligands are involved, and many of these standard methodologies fail when designed inhibitors with high binding constants and slow exchange rates are used. This collaborative project provides additional orientational constraints using new NMR methodology that measures residual dipolar couplings (RDCs), paramagnetic relaxation enhancements (PREs), and pseudo-contact shifts (PCSs) on ligands bound to the perdeuterated and lanthanide tagged carbohydrate recognition domain of galectin-3.