STRUCTURAL STUDIES ON THE PLASMODIUM SURFACE PROTEINS

在这项研究中，疟原虫寄生虫种类和结构 我们建议用来制备合成肽的阶段特异性表面蛋白 多肽链的不同部分的片段，包括串联 重复，所有疟疾疟原虫寄生虫表面的共同特征 到目前为止研究的蛋白质和合成的构象 与免疫优势串联重复序列(表位)相对应的肽使用 核磁共振和圆二色谱。这些合成纤维的表面性质 表位及其合成聚合物将在 单层实验。 我们还将设计重复表位的多肽类似物，并 其他多肽片段，试图提高高滴度和高亲和力 可能有效地开发出针对这些疾病的疫苗的抗体 疟疾寄生虫。诺氏疟原虫环子孢子蛋白将是 酵母和大肠杆菌中的表达及高效液体纯化 利用抗体定向的层析和亲和层析 对抗合成肽以及抗子孢子提取物。这个 将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)，并 用圆二色谱和核磁共振对其构象进行分析。 最后，我们将尝试使用合成材料来开发疟疾疫苗。 与CS蛋白片段相对应的多肽或其类似物。

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.