Proteinase Systems in Insect Hemolymph

昆虫血淋巴中的蛋白酶系统

• 批准号: 7940326
• 负责人: Michael R Kanost
• 金额: $ 25.77万
• 依托单位: KANSAS STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-30 至 2010-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7940326
• 关键词: Affect; Animals; Area; Bacteria; Binding; Biochemical; Biological Assay; Biological Models; Blood; Blood coagulation; Cell membrane; Cleaved cell; Clip; Complement; Complex; Culicidae; Deposition; Enzyme Precursors; Enzymes; Genetic; Genomics; Goals; Hemolymph; Hemorrhage; Immune response; Infection; Insect Vectors; Insecta; Invaded; Invertebrates; Investigation; Knowledge; Laboratories; Location; Manduca; Manduca sexta; Mediating; Melanins; Microbe; Modeling; Monophenol Monooxygenase; Organism; Outcome; Parasites; Pathway interactions; Pattern Recognition; Peptide Hydrolases; Peptides; Phospholipids; Plasma; Plasmodium; Polysaccharides; Preparation; Protease Domain; Protein Precursors; Proteins; Proteolytic Processing; Quinones; Regulation; Research; Research Personnel; Role; Serine Protease; Serpins; Signal Transduction; Site; Surface; System; Testing; Travel; Work; Wound Infection; base; cofactor; cytokine; design; extracellular; human disease; inhibitor/antagonist; killings; microbial; microorganism; pathogen; preference; programs; protein complex; research study; response; wound

Extracellular serine protease cascades modulate protective responses to limit bleeding and infection in vertebrate and invertebrate animals, but our understanding of these pathways in insects is rudimentary at best. Previous work from this laboratory has established the lepidopteran insect, Manduca sexta, as a model system well suited for biochemical characterization of the complex functions of hemolymph proteases. Experiments in this proposal are designed to investigate the roles of plasma serine proteases and their inhibitors (serpins) in insect innate immune responses, including activation of phenoloxidase. The work will test hypotheses based on the following model. Pattern recognition proteins in plasma bind to microbial surfaces. These proteins then interact with serine protease zymogens and other protein cofactors to form protein complexes, which localize the protease cascade and subsequent melanin deposition to the foreign surface. Separate pathways (or separate branches of converging pathways) may be initiated by different types of microorganisms, as they are recognized by separate sets of pattern recognition proteins. Inhibition of the proteases by specific interactions with serpins limit the duration and location of the response. The specific aims of the projectare: 1. Investigate the activation of selected clip domain proteases and their inhibition by specific serpins, to determine their order in microbe-activated cascade pathways. 2. Identify protein interactions that result in formation of protease activation complexes and investigate the assembly of such complexes and their roles in localizing phenoloxidase activation on surfaces. Relevance: Innate immune responses are likely to affect the outcome of infections of insect vectors with the pathogens and parasites they transmit. The long term goals of the research are to gain a thorough understanding of the protease cascades that mediate innate immune responses in M. sexta, to apply this knowledge to insect vectors of human diseases, and to apply advantages of the M. sexta system for fundamental studies on the regulation of serine protease activity.

细胞外丝氨酸蛋白酶级联调节保护性反应，以限制出血和感染， 脊椎动物和无脊椎动物，但我们对昆虫中这些途径的理解是初步的， 最好这个实验室以前的工作已经建立了鳞翅目昆虫，天蛾，作为一个模型， 系统非常适合血淋巴蛋白酶的复杂功能的生化表征。 本实验旨在研究血浆丝氨酸蛋白酶及其活性在糖尿病发病中的作用。 抑制剂（丝氨酸蛋白酶抑制剂）在昆虫先天免疫反应，包括激活酚氧化酶。这项工作将 根据以下模型检验假设。血浆中的模式识别蛋白与微生物结合 表面。然后这些蛋白质与丝氨酸蛋白酶酶原和其他蛋白质辅因子相互作用， 蛋白质复合物，其将蛋白酶级联和随后的黑色素沉积定位于外源性黑色素。 面单独的途径（或会聚途径的单独分支）可以由不同的 微生物类型，因为它们被不同的模式识别蛋白识别。抑制 蛋白酶通过与丝氨酸蛋白酶抑制剂的特异性相互作用而产生的反应限制了反应的持续时间和位置。 该项目的具体目标是： 1.研究选定的剪切结构域蛋白酶的激活及其被特异性丝氨酸蛋白酶抑制剂抑制， 确定它们在微生物激活级联途径中的顺序。 2.鉴定导致蛋白酶活化复合物形成的蛋白质相互作用，并研究蛋白质相互作用的机制。 这些复合物的组装及其在表面上定位酚氧化酶活化中的作用。 相关性：先天免疫反应可能影响昆虫媒介感染的结果， 它们传播的病原体和寄生虫。研究的长期目标是获得一个彻底的 了解介导M. sexta，应用这个 利用M.六进制 丝氨酸蛋白酶活性调节的基础研究。