STRUCTURAL STUDIES ON THE PLASMODIUM SURFACE PROTEINS

在这项对疟原虫疟疾寄生虫物种结构的研究中 我们建议制备合成肽的阶段特异性表面蛋白 多肽链各部分的片段，包括串联 重复，所有疟疾疟原虫寄生虫表面的共同特征 迄今为止研究的蛋白质并研究合成的构象 使用与免疫显性串联重复序列（表位）相对应的肽 NMR 和圆二色性。 这些合成材料的表面特性 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和 其他肽片段试图提高滴度和高亲和力 可能有效开发针对这些疾病的疫苗的抗体 疟疾寄生虫。 P. Knowlesi 环子孢子蛋白将是 在酵母和大肠杆菌中表达并通过高性能液体纯化 使用抗体定向的层析和亲和层析 对抗合成肽以及子孢子提取物。 的 将分析表达蛋白质的一级和二级结构， 两个可能的二硫桥（如果存在）将被确定并且 其构象将通过CD和NMR进行分析。 最后，将尝试使用合成材料开发疟疾疫苗 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.