REDUCTIVE CLEAVAGE METHOD FOR GLYCAN STRUCTURAL ANALYSIS

复杂碳水化合物结构的测定是一个重要的 在许多生物医学研究领域的问题，因为他们 参与细胞识别过程，它们作为结构 材料，以及它们在肿瘤中作为抗原决定簇的存在， 细菌真菌和病毒 由于其结构的复杂性， 建立它们结构的单一方法是可用的。 的无数 目前可用的化学方法是非常费力和模糊的 结果往往会得到。 因此，我们开始了一项计划， 用于复杂聚糖结构表征的新方法， 将克服与当前方法有关的困难， 和含有中性醛糖的聚糖的初步研究， 已经确定已经开发了这种方法。 构成这一建议基础的结构方法是指 称为"还原裂解法"，因为其显著特点是 区域特异性还原裂解的所有糖苷键在一个完全 甲基化多糖 这种方法与 "甲基化分析"，但是，因为1）所有的连锁位置和环 同时建立多糖中的形式，以及2）转化 将完全甲基化的多糖转化为适用于 g.l.c. - M.S.分析在一锅反应中完成， 中间转移。 根据我们最初获得的结果， 通过调查，我们认为还原裂解法具有 作为结构表征的一般方法的潜力 所有复杂的碳水化合物，和实验设计，以检查 这一做法的可行性构成了这一建议的基础。 研究 这里提出的也作为其目标的一体化的还原 裂解方法和以前开发的方法， 复杂碳水化合物结构分析的简化方案。

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 the current methodologies, and preliminary studies with glycans that contain neutral aldoses and ketoses have established that such a method has been developed. 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 from "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 g.l.c.-m.s. analysis is accomplished in a one-pot reaction without intermediate transfers. Based upon the results obtained from our initial investigations, we feel that the Reductive Cleavage Method has the potential to serve as a general method for the structural characterization of all complex carbohydrates, and experiments designed to examine the feasibility of this approach form the basis of this proposal. The research proposed here also has as its objective the integration of the Reductive Cleavage Method and previously-developed methodologies into an overall, simplified scheme for complex carbohydrate structural analysis.