Mass Spectrometry of Metal-Coordinated Oligosaccharides

描述(由申请人提供)：本次竞争更新的主要目标集中在糖胺多糖的分离、纯化和完整的化学表征上，因为它与肝素和HS对各种炎性趋化因子的络合作用有关。建立了三种特异性类风湿关节炎相关趋化因子的糖胺多聚糖(GAG)结合体的分离、纯化和鉴定方法，并用离子迁移质谱(IM-MS)和CID-MS进行鉴定。GAG结合体与其对应的趋化因子的络合作用将用MS进行研究，单体、二聚体和四聚体组装用IM-MS测量。这些研究将与绝经前和绝经后年龄范围的类风湿性关节炎妇女血清中GAG的成分分析相结合，以确定HRT的健康益处及其与GAG成分的相关性。这三个特异靶是：特异靶1：人血清中的GAG组成，特异靶2：GAG文库与趋化因子的色谱制备、分离、鉴定和络合，以及特异靶3：趋化因子和GAG结合物的离子迁移率MS和X射线结晶学。建议的主要领域以这样一种方式交织在一起，即方法开发、生物技术和对生物相关系统的应用是将建议的研究捆绑在一起的紧密联系的要素。

DESCRIPTION (provided by applicant): The broad goal of this competing renewal focuses on glycosaminoglycan separation, purification and complete chemical characterization as it pertains to heparin and HS complexation to various inflammatory chemokines. Methods will be developed to separate, purify and identify glycosaminoglycan (GAG) binders of three specific Rheumatoid Arthritis associated chemokines using chromatography, ion mobility mass spectrometry (IM-MS) and CID-MS. Complexation of GAG binders to their chemokine counterparts will be investigated by MS with the monomer, dimer and tetramer assemblies measured using IM-MS. These studies will be paired with the compositional analysis of GAG in sera from women with Rheumatoid Arthritis in the pre- and postmenopausal age range to determine health benefits of HRT and its correlation with GAG composition. The three specific aims are: Specific Aim 1: GAG composition in Human Serum, Specific Aim 2: Chromatographic preparation, separation, identification and complexation of GAG libraries with Chemokines, and Specific Aim 3: Ion Mobility MS and X-ray crystallography of Chemokine and GAG binders. 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.