REDUCTIVE CLEAVAGE METHOD FOR GLYCAN STRUCTURAL ANALYSIS

复杂碳水化合物结构的测定是一种 在许多生物医学研究领域中的重要问题，因为 它们在细胞识别过程中的参与，它们的作用 作为结构材料，以及作为抗原性的存在 肿瘤、细菌、真菌和病毒中的决定因素。因为. 它们的结构复杂性，没有单一的方法来建立 他们的结构是可用的。无数化学方法论 目前可用的是非常费力和模棱两可的结果 通常是获得的。因此，我们开始了一项计划，开发一种新的 一种用于表征具有以下结构的复合多糖的方法 将克服与当前 方法论。 构成这项建议基础的结构方法是 被称为“约化割裂法”是因为它的显著特点 特征是所有糖苷的区域特异性还原裂解 完全甲基化的多糖中的键。这种方法是 然而，与“甲基化分析”本质不同的是， 因为1)多糖中的所有连接位置和环形式 是同时建立的，以及2)完全转换 适合分离的甲基化多糖衍生物 而分析是在一部过去的作品中完成的，我们觉得 约化割裂方法具有作为一般方法的潜力 所有络合物的结构表征方法 碳水化合物，包括确定 组成单体，它们的环形式，它们的位置(S) 连锁、它们的异常构型和它们的序列。 为检验这种方法的可行性而设计的实验 形成了这项建议的基础。在此提出的研究 因此，它以还原的整合为目标 裂解法解复杂碳水化合物的简化方案 结构分析。

The determination of structure of complex carbohydrates is an important problem in many areas of biomedical investigation because of their involvement in cellular recognition processes, their role as structural materials, and their presence as antigenic determinants in tumors, bacteria, fungi, and viruses. Because of their structural complexity, no single method for establishing their structure is available. The myriad of chemical methodologies currently available are very laborious and ambiguous results are often obtained. We therefore began a program to develop a new method for the structural characterization of complex glycans that would overcome the difficulties associated with current methodologies. The structural method that forms the basis of this proposal is referred to as the "Reductive Cleavage Method" because its salient feature is the regiospecific reductive cleavage of all glycosidic bonds in a fully methylated polysaccharide. This method is substantially different than "methylation analysis", however, because 1) all linkage positions and ring forms in a polysaccharide are established simultaneously, and 2) conversion of the fully methylated polysaccharide to derivatives suitable for separation and analysis is accomplished in a one-past work, we feel that the Reductive Cleavage Method has the potential to serve as a general method for the structural characterization of all complex carbohydrates, to include establishing the identities of constituent monomers, their ring form, their position(s) of linkage, their anomeric configuration, and their sequence. Experiments designed to examine the feasibility of this approach form the basis of this proposal. The research proposed herein therefore has as its objectives the integration of the Reductive Cleavage Method into a simplified scheme for complex carbohydrate structural analysis.