Mass Spectrometry of Metal-Coordinated Oligosaccharides

这一竞争性更新的广泛目标集中在研究 碳水化合物：蛋白质，以及蛋白质：蛋白质非共价相互作用。特别是， 我们建议研究糖胺聚糖（GAG）与 N-末端G-蛋白偶联受体（GPCR）的趋化因子配体，CCR 2和 CCR 5，以及使用Q-TOF/Ion探索所有三种可能的三元复合物 迁移率质谱法。将通过以下方法探索GAG与趋化因子的结合： 将各种GAG文库与趋化因子CCR 2和CCR 5孵育。具体 将使用我们的GAG组成分析鉴定粘合剂。趋化因子将是 在室内进行表达和纯化。将进行碰撞截面测量 对于所鉴定的任何非共价复合物，从而提供关于 结合时构象发生变化。额外的表面等离子体共振和MS 结合动力学研究沿着热量测定法将提供重要的信息 关于结合强度。目的是详细了解这些 相互作用将允许有效靶向趋化因子系统， 效益所提出的主要领域是相互交织的， 发展，生物技术和生物相关系统的应用是 将拟议的研究结合在一起的有凝聚力的元素。

The broad goal of this competing renewal focuses on studying carbohydrate:protein, as well as protein:protein non-covalent interactions. In particular, we propose to investigate the binding and interaction of glycosaminoglycans (GAG) with chemokine ligands of the N-terminal G-protein coupled receptors (GPCR), CCR2 and CCR5, as well as exploring possible ternary complexes of all three using Q-TOF/Ion Mobility Mass Spectrometry. Binding of the GAG with chemokine will be explored via incubation of various GAG libraries with the chemokines of CCR2 and CCR5. Specific binders will be identified using our GAG compositional analysis. Chemokines will be expressed and purified in house. Collisional cross section measurements will be made for any of the non-covalent complexes identified, thus providing information about conformation change upon binding. Additional Surface Plasmon resonance and MS kinetic binding studies along with thermal calorimetry will provide important information about the binding strength. The intent is that a detailed understanding of these interactions will allow for effective targeting of the chemokine system for therapeutic benefit. The major areas proposed are interwoven in such a way that methods development, biotechnology and application to biologically relevant systems are the cohesive elements that bind the proposed research together.