VACUOLAR & MEMBRANES OF ISOLATED PLASMODIUM

建议对毛白杨的比较关键性能进行评价。 诺氏，P，k和恶性疟原虫，P.F，空泡膜，PVM， (A)继续使用无宿主细胞分离的P.K裂殖体进行工作 膜污染由多个标准确定 P.K PVM的重要结构和功能属性 蛋白质，以及(B)将我们的方法逐步纳入一项研究(已经 已启动)到P.F PVM。 计划采取以下方法： 1.猪瘟病毒外表面暴露的宿主细胞蛋白质分析 PVM通过载体表面标记，选择性多肽切割， 和使用特定的抗体，并特别注意 (A)红细胞蛋白3，(B)血糖素、糖蛋白和(C)血红蛋白。 2.PVM中寄生虫合成蛋白的检测，使用 代谢标记与载体表面标记相结合， 选择性多肽切割和特定抗体，与特定的 关注我们先前已知的74 kDa P.K蛋白 在P.KPVM中检测到，可能与P.F. 3.功能研究(A)测量阴离子通过 (B)使用阴离子通道探针评价PVM的作用 在这一过程中，(C)评估外源ATP的能力 支持分离的裂殖体的蛋白质合成，(D)使用 光亲和探针定位苍术皂苷的路线- 敏感的三磷酸腺苷进入。 4.开发用于(A)选择性增溶PVM的方法， 和(B)将PVM蛋白插入磷脂双层，以 纯化完整的PVM蛋白，允许更广泛的未来研究 在PVM上，并作为最终探索膜的手段 红内型疟原虫的交通。

It is proposed to evaluate the compare key properties of P. knowlesi, P,k, and P falciparum, P.f, vacuolar membranes, PVMs, (a) continuing work using P.k schizonts, isolated free of host cell membrane contamination by multiple criteria, to determine important structural and functional attributes of P.k PVM proteins, and (b) phasing our approaches into a study (already initiated) to the P.f PVM. The following approaches are planned: 1. Analysis of host cell proteins exposed at he outer surface of the PVM by vectorial surface labeling, selective peptide cleavage, and use of specific antibodies, with specific attention directed to (a) erythrocyte protein 3, (b) glycophorin, GP, and (c) hemoglobin. 2. Examination of parasite-synthesized proteins in the PVM, using metabolic labeling combined with vectorial surface labeling, selective peptide cleavage, and specific antibodies, with specific attention on the 74 kDa P.k protein which we have previously detected in the P.k PVM, and possibly related proteins of P.f. 3. Functional studies (a) measuring anion transport through the PVM, (b) using anion channel probes to evaluate the role of PVM in this process, (c) evaluating the ability of exogenous ATP to support protein synthesis of isolated schizonts, (d) using photoaffinity probes to localize the route of atractyloside- sensitive ATP entry. 4. Develop methods for (a) selective solubilization of the PVM, and (b) the insertion of PVM proteins into phospholipid bilayers, to purify integral PVM proteins, allow more extensive future studies on the PVM, and as means for eventual exploration of membrane traffic in intraerythrocytic Plasmodium.