REDUCTIVE CLEAVAGE METHOD FOR GLYCAN STRUCTURAL ANALYSIS

复杂碳水化合物的结构测定是一个重要的研究课题 生物医学研究的许多领域中的问题，因为他们的 参与细胞识别过程，它们在结构上的作用 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。由于它们的结构复杂性，没有 建立其结构的单一方法是可用的。无数的 目前可用的化学方法非常费力且模棱两可。 结果往往是可以得到的。因此，我们开始了一项计划，开发一种 一种表征复杂多聚糖结构的新方法 将克服与当前方法相关的困难， 以及对含有中性醛糖和糖链的多糖的初步研究 酮糖已经证实，这种方法已经开发出来。 参考构成本提案基础的结构方法 因为它的显著特征是 所有糖苷键的区域特异性还原裂解 甲基化多糖。这种方法与 然而，“甲基化分析”，因为1)所有的连接位置和环 同时建立多糖中的各种形式，并2)转化 将完全甲基化的多糖转化为适合于 G.L.C.-M.S.分析是在一锅反应中完成的，没有 中间转账。根据我们从初步调查中获得的结果 调查中，我们认为还原割裂法具有 有可能作为结构表征的一般方法 所有复杂的碳水化合物，以及旨在研究 这一方法的可行性构成了这项建议的基础。这项研究 这里提出的也以还原的积分作为其目标 将分解方法和以前开发的方法整合为一个整体， 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.