Function and regulation of two key serine proteases in insect immunity

两种关键丝氨酸蛋白酶在昆虫免疫中的功能和调控

• 批准号: 10246410
• 负责人: HAOBO JIANG
• 金额: $ 34.55万
• 依托单位: OKLAHOMA STATE UNIVERSITY STILLWATER
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-03-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10246410
• 关键词: Antibodies; Area; Bacteria; Binding Proteins; Biochemical; Biological Assay; Chemicals; Clip; Collection; Complex; Constitution; Culicidae; Data; Defense Mechanisms; Detection; Disease Vectors; Drosophila genus; Enzyme Precursors; Evolution; Fractionation; Future; Genes; Genetic; Genome; Glucans; Goals; Gram-Negative Bacteria; Gram-Positive Bacteria; Hemolymph; Histidine; Immune; Immune response; Immune signaling; Immunity; Infection; Innate Immune System; Insect Vectors; Insecta; Invaded; Investigation; Knowledge; Larva; Lead; Ligands; Malaria; Manduca; Manduca sexta; Mass Spectrum Analysis; Mediating; Melanins; Microbe; Modeling; Molecular; Molecular Probes; Monophenol Monooxygenase; Natural Immunity; Organism; Orthologous Gene; Parasites; Pathway interactions; Peptide Hydrolases; Peptides; Peptidoglycan; Plasma; Play; Process; Production; Property; Protein Biosynthesis; Proteins; Recombinant Proteins; Regulation; Research; Role; Serine Protease; Signal Pathway; Signal Transduction; Specificity; Stress; Study models; System; Testing; antimicrobial; antimicrobial peptide; base; biochemical model; cytokine; defense response; experience; extracellular; fungus; human disease; improved; innate immune pathways; insect disease vector; insight; pathogen; peptidoglycan recognition protein; public health relevance; response; thioester; transmission process; vector

Project Summary Extracellular serine proteases and their non-catalytic homologs coordinate key defense mechanisms in insects. This includes responses of mosquitoes against pathogens and parasites that cause serious human diseases. Proteolytic cleavage generates active phenoloxidases (POs), thioester proteins (TEPs), Spätzle and other cytokines. PO catalyzes the production of reactive chemicals to sequester and kill the invading organisms. TEPs opsonize parasites, targeting them for destruction. Spätzle and other cytokines trigger intracellular signaling pathways to induce the expression of antimicrobial peptides and other defense proteins. In insect vectors of human diseases, the protease networks may be evaded or disrupted by proteins from the intruders. Knowledge of the system components and their interactions from biochemical model insects such as Manduca sexta is useful for gaining detailed molecular understanding of fundamental aspects of insect innate immunity, including these protease cascades and provides basic knowledge that can guide studies of similar systems in insect disease vectors. We have elucidated a part of the M. sexta protease network that activates proPO in response to bacteria and fungi. In this network, recognition proteins bind to microbes and activate the proteases in a cascade mode. We have annotated 193 genes encoding serine proteases and their homologs, identified 36 of the proteins in larval hemolymph, and acquired data indicating that some of them play critical roles in immune signal transduction. Based on the molecular probes, purified proteins and working experience, we propose to investigate two critical steps of the protease network, which are conserved in dipteran species, by combining hemolymph fractionation, recombinant protein production and processing, sequence and expression information, and state-of-the-art mass spectrometry. The specific aims of this project are to: 1) characterize the system initiation by examining interactions of peptidoglycans, two recognition proteins, and hemolymph protease-14 precursor (proHP14) at the domain level; 2) identify 1−2 proHP6-activating proteases and elucidate their activation mechanisms. New knowledge gained in this project will improve understanding of insect immunity at a biochemical level and stimulate related research in vector species.

项目摘要 细胞外连续蛋白酶及其非催化同源物协调关键防御机制 在昆虫中。这包括蚊子对病原体和寄生虫的反应 严重的人类疾病。蛋白水解裂解会产生活性苯酚酶（POS），硫酯 蛋白质（TEP），Spätzle和其他细胞因子。 PO催化反应性化学物质的产生 隔离并杀死入侵的生物。 teps对寄生虫进行调整，以销毁为目标。 Spätzle和其他细胞因子触发细胞内信号通路诱导的表达 抗菌胡椒和其他防御蛋白。在人类疾病的昆虫载体中，蛋白质 网络可能会因入侵者的蛋白质而逃避或破坏。系统知识 来自生化模型昆虫（如甘达·塞克斯塔）的成分及其相互作用很有用 为了获得对昆虫先天免疫基本方面的详细分子理解， 包括这些蛋白酶级联 昆虫疾病向量的系统。我们已经阐明了M. sexta蛋白酶网络的一部分 激活对细菌和真菌的响应。在此网络中，识别蛋白结合到 微生物并在级联模式下激活蛋白酶。我们已经注释了193个编码的基因 连续蛋白及其同源物，在幼虫血淋巴中鉴定出36个蛋白质，并获得 数据表明其中一些在免疫信号传递中起关键作用。基于 分子问题，纯化的蛋白质和工作经验，我们建议研究两个关键 蛋白酶网络的步骤通过结合血淋巴在双翅目物种中保守的步骤 分馏，重组蛋白的产生和加工，序列和表达信息， 和最先进的质谱。该项目的具体目的是：1）表征 通过检查胡椒糖的相互作用，两个识别蛋白和 在域水平上的血淋巴蛋白酶14前体（ProHP14）； 2）确定1-2个proHP6激活 蛋白酶并阐明其激活机制。这个项目中获得的新知识将 在生化水平上提高对绝缘免疫力的理解，并刺激相关研究 向量物种。

项目成果

Comparative genomic analysis of the Tribolium immune system.

• DOI: 10.1186/gb-2007-8-8-r177
• 发表时间: 2007
• 期刊: Genome biology
• 影响因子: 12.3
• 作者: Zou Z;Evans JD;Lu Z;Zhao P;Williams M;Sumathipala N;Hetru C;Hultmark D;Jiang H
• 通讯作者: Jiang H

A mechanistic analysis of bacterial recognition and serine protease cascade initiation in larval hemolymph of Manduca sexta.

• DOI: 10.1016/j.ibmb.2022.103818
• 发表时间: 2022-09
• 期刊: INSECT BIOCHEMISTRY AND MOLECULAR BIOLOGY
• 影响因子: 3.8
• 作者: Wang, Yang;Kanost, Michael R.;Jiang, Haobo
• 通讯作者: Jiang, Haobo

Nitric Oxide-Induced Calcineurin A Mediates Antimicrobial Peptide Production Through the IMD Pathway.

一氧化氮诱导的钙调神经磷酸酶 A 通过 IMD 途径介导抗菌肽的产生

• DOI: 10.3389/fimmu.2022.905419
• 发表时间: 2022
• 期刊: FRONTIERS IN IMMUNOLOGY
• 影响因子: 7.3
• 作者: Chen, Kangkang;Wang, Xinyan;Wei, Xiangyi;Chen, Jiaqian;Wei, Youheng;Jiang, Haobo;Lu, Zhiqiang;Feng, Congjing
• 通讯作者: Feng, Congjing

A comprehensive analysis of the Manduca sexta immunotranscriptome.

• DOI: 10.1016/j.dci.2012.10.004
• 发表时间: 2013-04
• 期刊: DEVELOPMENTAL AND COMPARATIVE IMMUNOLOGY
• 影响因子: 2.9
• 作者: Gunaratna, Ramesh T.;Jiang, Haobo
• 通讯作者: Jiang, Haobo

The Micrococcus luteus infection activates a novel melanization pathway of cSP10, cSP4, and cSP8 in Helicoverpa armigera.

• DOI: 10.1016/j.ibmb.2022.103775
• 发表时间: 2022-04
• 期刊: Insect biochemistry and molecular biology
• 影响因子: 3.8
• 作者: Qianran Wang;Mengyi Yin;Chuanfei Yuan;Xi-Juan Liu;Haobo Jiang;Manli Wang;Z. Zou;Zhìhóng Hú