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.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM041247-21
Application #
7746467
Study Section
Vector Biology Study Section (VB)
Program Officer
Jones, Warren
Project Start
1991-07-01
Project End
2011-04-04
Budget Start
2010-01-01
Budget End
2011-04-04
Support Year
21
Fiscal Year
2010
Total Cost
$312,732
Indirect Cost
Name
Kansas State University
Department
Biochemistry
Type
Schools of Arts and Sciences
DUNS #
929773554
City
Manhattan
State
KS
Country
United States
Zip Code
66506
Al Souhail, Qasim; Hiromasa, Yasuaki; Rahnamaeian, Mohammad et al. (2016) Characterization and regulation of expression of an antifungal peptide from hemolymph of an insect, Manduca sexta. Dev Comp Immunol 61:258-68
Kanost, Michael R; Arrese, Estela L; Cao, Xiaolong et al. (2016) Multifaceted biological insights from a draft genome sequence of the tobacco hornworm moth, Manduca sexta. Insect Biochem Mol Biol 76:118-147
Cao, Xiaolong; He, Yan; Hu, Yingxia et al. (2015) Sequence conservation, phylogenetic relationships, and expression profiles of nondigestive serine proteases and serine protease homologs in Manduca sexta. Insect Biochem Mol Biol 62:51-63
Cao, Xiaolong; Jiang, Haobo (2015) Integrated modeling of protein-coding genes in the Manduca sexta genome using RNA-Seq data from the biochemical model insect. Insect Biochem Mol Biol 62:2-10
Takahashi, Daisuke; Garcia, Brandon L; Kanost, Michael R (2015) Initiating protease with modular domains interacts with ?-glucan recognition protein to trigger innate immune response in insects. Proc Natl Acad Sci U S A 112:13856-61
Koenig, Christopher; Hirsh, Ariana; Bucks, Sascha et al. (2015) A reference gene set for chemosensory receptor genes of Manduca sexta. Insect Biochem Mol Biol 66:51-63
Kanost, Michael R; Jiang, Haobo (2015) Clip-domain serine proteases as immune factors in insect hemolymph. Curr Opin Insect Sci 11:47-55
Peng, Zeyu; Dittmer, Neal T; Lang, Minglin et al. (2015) Multicopper oxidase-1 orthologs from diverse insect species have ascorbate oxidase activity. Insect Biochem Mol Biol 59:58-71
He, Yan; Cao, Xiaolong; Li, Kai et al. (2015) A genome-wide analysis of antimicrobial effector genes and their transcription patterns in Manduca sexta. Insect Biochem Mol Biol 62:23-37
Zhang, Xiufeng; He, Yan; Cao, Xiaolong et al. (2015) Phylogenetic analysis and expression profiling of the pattern recognition receptors: Insights into molecular recognition of invading pathogens in Manduca sexta. Insect Biochem Mol Biol 62:38-50

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