PHYSIOLOGY OF INSECT AMINO ACID TRANSPORT

昆虫氨基酸运输的生理学

• 批准号: 6169630
• 负责人: WILLIAM R HARVEY
• 金额: $ 22.65万
• 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/6169630
• 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; 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：碱金属离子分类蛋白 甘达·塞克斯塔（Manduca Sexta）碱性中肠和埃及埃及埃及的蛋白质，到 分析共异体在化学偶联氨基酸中的作用 吸收并开发这些独特转运蛋白的抑制剂 环境安全的蚊子。 假设是独特的 毛毛虫和幼虫蚊子中的碱性中肠含量反映了 质膜的独特营养摄取方法 通过H+易位，液泡型ATPase和A燃烧，有氧运动。 k+/2h+抗胞毒剂。 电动膜上的电压驱动氨基 酸：K+交构入细胞。 独特的K+：氨基酸转运蛋白， KAAT1，最近从M. Sexta Midgut和其他Caterpillar克隆 推测转运蛋白介导蚊子幼虫中的氨基酸摄取 也是如此。 目的1是分析KAAT1及其同工型的作用机理。 Sexta和Ae。在放松动力学和现场定向方面 阳离子和氨基酸结合位点的诱变。 目标2是克隆 并表征编码系统B，Pro-Gly和R+的cDNA和M. Sexta和R+的R+ Ae。埃及。 AIM 3是将KAAT1和其他转运蛋白定位在M. Sexta中 和Ae。埃及细胞通过原位杂交和免疫细胞化学。 目的 4是在平面脂质双层中重建KAAT1和其他对称器 研究特定膜脂质对动力学的影响 特征。 AIM 5，与商业公司（Rohm和Haas）一起 确定KAAT1和其他昆虫转运蛋白的特定抑制剂，并 将它们作为环境安全的幼虫发展。 该项目有科学 优点是因为像KAAT1一样，其他转运蛋白可能是独一无二的， 比na+耦合的K+ frather是由电压驱动的 在体内pH 9.5和11.5之间运行。 很可能是 KAAT1和这些推定的转运蛋白可以作为安全的蚊子开发 只能在高肠道pH下笼和有选择地激活的幼虫。