LEISHMANIA PROTON PUMP: GENES & PROTEINS

L.主动运输donovani依赖于质子梯度 穿过细胞膜， 一个能挤出质子的ATP酶。 这个质子还必须扮演一个 在酸性环境中寄生虫的生存至关重要的作用 环境的脊椎动物宿主的吞噬溶酶体。 我们的长期 目的是了解结构，功能和调节 这种酶，并通过这种理解提供途径， 控制这种重要的病原体。 这项建议的目的是 是应用分子克隆技术， 用于分析介导细胞凋亡的大分子的探针 质子泵系统 我们在第一阶段取得了成功， 并克隆了两个串联基因的一部分 在结构上能够编码质膜阳离子 泵. 我们将克隆这对串联的L。donovani基因在 他们的全部 我们将确定基因的DNA序列， 并预测其蛋白质产物的氨基酸序列。 该序列信息将用于生产核酸和 用于分析两种生命中的表达的抗体探针 寄生虫的周期阶段。 我们将比较 这些基因产物的质子泵活性，并检查 推测的蛋白质编码的酶活性 质子泵基因

Active transport in L. donovani is dependent on a proton gradient across the plasma membrane that is established through the action of an ATPase that extrudes protons. This proton must also play a vital role in the survival of the parasite within the acidic environment of the vertebrate host's phagolysosomes. Our long term goal is to understand the structure, function and regulation of this enzyme and, through this understanding to provide avenues to the control of this important pathogen. The aim of this proposal is to apply molecular cloning technology to produce molecular probes with which to analyze the macromolecules that mediate the proton pumping system. We have succeeded in the first phase of this endeavor and have cloned parts of two tandemly arrayed genes that are structurally capable of encoding plasma membrane cation pumps. We will clone this tandem pair of L. donovani genes in their entirety. We will determine the DNA sequences of the genes, and predict the amino acid sequences of their protein products. This sequence information will be used produce nucleic acid and antibody probes for the analysis of expression in the two life cycle stages of the parasite. We will compare the abundance of these gene products to proton pumping activity, and examine the enzymatic activities of the proteins encoded by the presumptive proton pump genes.