STRUCTURAL STUDIES ON THE PLASMODIUM SURFACE PROTEINS

在这项研究中，疟原虫疟疾寄生虫种类的结构和 阶段特异性表面蛋白我们建议制备合成肽 多肽链的各个部分的片段，包括串联的 重复，这是所有疟疾疟原虫表面的共同特征 蛋白质研究的日期和研究的构象的合成 对应于免疫显性串联重复序列（表位）的肽， NMR和圆二色性。 这些合成材料的表面性质 表位及其合成制备的聚合物将在 单层实验 我们还将设计串联重复表位的肽类似物， 其它肽段，以试图提高高滴度和高亲和力 这些抗体可以有效地开发针对这些疾病的疫苗， 疟原虫 诺氏毕赤酵母环子孢子蛋白将被 在酵母和E.大肠杆菌，并经高效液相色谱纯化 使用抗体定向的层析和亲和层析 针对合成肽以及针对子孢子提取物。 的 分析表达蛋白的一级和二级结构， 将确定两个可能的二硫键（如果存在的话）， 用圆二色谱和核磁共振对其构象进行分析。 最后，将尝试使用合成的 对应于CS蛋白片段的肽或其类似物。

In this study of the structure of Plasmodium malaria parasite species and stage-specific surface proteins we propose to prepare synthetic peptide segments of various parts of the polypeptide chain including the tandem repeats, a common feature of all malaria Plasmodium parasite surface proteins studied to date and study the conformation of the synthetic peptide corresponding to the immunodominant tandem repeats (epitopes) using NMR and circular dichroism. The surface properties of these synthetic epitopes and their synthetically prepared polymers will be measured in monolayer experiments. We shall also design peptide analogs of the tandemly repeated epitopes and other peptide segments in an attempt to raise high titer and high affinity antibodies which may be effective in developing a vaccine against these malaria parasites. The P. Knowlesi circumsporozoite protein will be expressed in yeast and E. coli and purified by high performance liquid chromatography and affinity chromatography using antibodies directed against the synthetic peptides as well as against sporozoite extracts. The primary and secondary structure of the expressed protein will be analyzed, the two possible disulfide bridges, if they exist, will be determined and its conformation will be analyzed by CD and NMR. Finally, an attempt will be made to develop malaria vaccine using synthetic peptides or their analogs corresponding to segments of the CS protein.