PHYSIOLOGY OF INSECT AMINO ACID TRANSPORT

昆虫氨基酸运输的生理学

• 批准号: 2628753
• 负责人: WILLIAM R HARVEY
• 金额: $ 25.14万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 1990
• 资助国家: 美国
• 起止时间: 1990-12-01 至 2003-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2628753
• 关键词: Aedes; Manduca; Xenopus oocyte; aminoacid transport; bioassay; brush border membrane; confocal scanning microscopy; disease vectors; homeostasis; immunocytochemistry; in situ hybridization; insect control; ion transport; larva; membrane transport proteins; molecular cloning; nucleic acid sequence; physiology; polymerase chain reaction; protein binding; protein structure function; site directed mutagenesis

DESCRIPTION (Adapted from the Applicant's Abstract): The long term objective is to clone and characterize amino acid:alkali metal ion symporter proteins from alkaline midguts of Manduca sexta and Aedes aegypti, to analyze the role of the symporters in chemiosmotically coupled amino acid uptake and to develop inhibitors of these unique transporters as environmentally safe mosquitocides. The hypothesis is that the uniquely alkaline midgut contents in caterpillars and larval mosquitoes reflect a unique method of nutrient uptake in which the plasma membranes are energized, aerobically, by an H+ translocating, vacuolar-type ATPase and a K+/2H+ antiporter. The voltage across the energized membrane drives amino acid:K+ symport into the cells. The unique K+:Amino Acid Transporter, KAAT1, recently cloned from M. sexta midgut, and other caterpillar transporters are postulated to mediate amino acid uptake in mosquito larvae as well. Aim 1 is to analyze the mechanism of action of KAAT1 and its isoforms in M. sexta and Ae. aegypti in terms of relaxation kinetics and site-directed mutagenesis of both cation and amino acid binding sites. Aim 2 is to clone and characterize cDNAs encoding Systems B, Pro-Gly, and R+ from M. sexta and Ae. aegypti. Aim 3 is to localize KAAT1 and other transporters in M. sexta and Ae. aegypti cells by in situ hybridization and immunocytochemistry. Aim 4 is to reconstitute KAAT1 and other symporters in planar lipid bilayers and to study the influence of specific membrane lipids on their kinetic characteristics. Aim 5, with a commercial firm (Rohm and Haas) is to identify specific inhibitors of KAAT1 and other insect transporters and to develop them as environmentally safe larvacides. The project has scientific merit because, like KAAT1, the other transporters are likely to be unique, to be K+ -rather than Na+ -coupled, to be driven by the voltage and to operate between pH 9.5 and 11.5 in vivo. It is likely that inhibitors of KAAT1 and these putative transporters may be developed as safe mosquito larvacides that can be caged and selectively activated only at high gut pH.

描述（改编自申请人的摘要）：长期 目的是克隆和表征氨基酸：碱金属离子同向转运体 来自天蛾和埃及伊蚊的碱性中肠的蛋白质， 分析同向转运蛋白在化学渗透偶联氨基酸中的作用 摄取并开发这些独特转运蛋白的抑制剂 对环境安全的杀蚊剂。 假设是唯一的 毛毛虫和蚊子幼虫的碱性中肠含量反映了 质膜吸收营养的独特方法 由 H+ 易位、液泡型 ATP 酶和有氧供能 K+/2H+ 反向转运蛋白。 通电膜上的电压驱动氨基 酸：K+转运进入细胞。 独特的K+：氨基酸转运蛋白， KAAT1，最近从 M. sexta 中肠和其他毛毛虫中克隆 假设转运蛋白介导蚊子幼虫的氨基酸摄取 以及。 目的 1 是分析 KAAT1 及其亚型在 M. 中的作用机制。 sexta 和 Ae。埃及伊蚊在弛豫动力学和位点定向方面 阳离子和氨基酸结合位点的诱变。 目标2是克隆 并表征来自 M. sexta 的编码系统 B、Pro-Gly 和 R+ 的 cDNA 和 艾。埃及伊蚊。 目标 3 是定位 M. sexta 中的 KAAT1 和其他转运蛋白 和艾。通过原位杂交和免疫细胞化学分析埃及细胞。 目的 4 是在平面脂质双层中重建 KAAT1 和其他同向转运蛋白 研究特定膜脂对其动力学的影响 特征。 与商业公司（罗门哈斯）合作的目标 5 是 鉴定 KAAT1 和其他昆虫转运蛋白的特异性抑制剂并 将它们开发为环境安全的杀幼虫剂。 该项目具有科学性 优点是因为，像 KAAT1 一样，其他转运蛋白可能是独一无二的， 是 K+ 而非 Na+ 耦合，由电压驱动并 体内 pH 值在 9.5 至 11.5 之间运行。 抑制剂很可能是 KAAT1 和这些假定的转运蛋白可能被开发为安全的蚊子 杀幼虫剂只能在高肠道 pH 值下被笼养和选择性激活。