Yang Wang, Senior Research Specialist
Native of Shanghai, China
1986 B.S., East China University of Chemical Technology, Biochemistry
1994 M.S., Kansas State University, Biochemistry
Zhen Zou, Postdoctoral Research Associate
Native of Shanghai, China
1997 B.S., East China University of Science and Technology, Biochemical engineering
2005 Ph.D., Oklahoma State University
Zhiqiang Lu, Graduate Research Assistant
Native of Shannxi, China
1994 B.S., Northwestern Agricultural University, Agronomy
1997 M.S., Northwestern Agricultural University, Biochemistry
Rudan Huang, Graduate Research Assistant
Native of Jiujiang, China
2004 B.S., University of Science and Technology of China, Biology
Rayaprolu Subrahmanyam, Graduate Research Assistant
Native of Kakinada, India
2000 B.S., Andhra University, Microbiology
2002 M.S., Andhra University, Microbiology
Picheng Zhao, Graduate Research Assistant
Native of Shanghai, China
2001 B.S., East China University of Science and Technology, Biochemical engineering
Siwei Liu, Graduate Research Assistant
Native of Henan, China
1983 B.S., Henan Agricultural University, Plant Protection
2005 M.S., Oklahoma State University, Entomology
Current research:
Extracellular serine proteinase cascades have evolved in vertebrates and invertebrates to mediate rapid defense reactions to wounding and infection. We have started to investigate such enzyme system and its regulation by proteinase inhibitors in a biochemical model insect, Manduca sexta. In collaboration with Dr. Kanost's group at Kansas State University, we are investigating the prophenoloxidase-activating pathway, a defense mechanism triggered by invading pathogens. Active phenoloxidase catalyzed the formation of reactive intermediates that are utilized to kill pathogens, seal wounds, and encapsulate parasites.
Prophenoloxidase is activated via limited proteolysis. We have purified three different prophenoloxidase-activating proteinases (PAPs) from M. sexta and examined their expression patterns. Transcription of these clip-domain serine proteinases is regulated in tissue-, development-, and defense-specific manners.
We have isolated cDNA clones for 22 different hemolymph proteinases, some of which are members of the serine proteinase system that coordinates the immune responses in M. sexta hemolymph. A modular protein, designated M. sexta HP14, interacts with ß-1,3-glucan or peptidoglycan, autoactivates, and triggers the prophenoloxidase-activating pathway.
This proteinase pathway is negatively regulated by serine proteinase inhibitors of the serpin superfamily. M. sexta serpin-1 variants, serpin-3, -4, -5, and -6 form stable proteinase-inhibitor complexes with one or more of the pathway components. Since molecular probes (e.g., cDNAs and antisera for HPs and serpins) are available, we are attempting to elucidate the prophenoloxidase-activating pathway, its relationships with other proteinase-mediated immune responses, and inhibitory regulation by serpins.
Laboratory Specialization:
hemolymph proteinases involved in insect immune responses
protein purification by HPLC/FPLC
characterization of hemolymph proteins
serine proteinase inhibitors of the serpin superfamily
expression of proteins in E. coli and baculovirus expression systems
molecular cloning of cDNAs and genes
transcriptional regulation of immune genes
Invited Symposium Presentations:
1996.05 International Symposium on the Chemistry and Biology of Serpins.
1996.07 Gordon Research Conference ("Proteolytic Enzymes and their Inhibitors")
2000.08 XXI International Congress of Entomology
2000.10 Annual Meeting of the Arbeitskreis Comparative Immunology, Germany
2003.07 Symposium on the Molecular Basis of the Recognition of Foreignness in Innate Immunity, Sapporo, Japan
2004.09 International Symposium on protein science in arthropods, Tsukuba, Japan
2005.12 The 4th Symposium of Biological and Medical Sciences at TMMU, Chongqing, China
Selected Publications:
1. Jiang, H., Wang, Y., and Kanost, M.R. (1994) Mutually exclusive exon use and reactive center diversity in insect serpins. J. Biol. Chem. 269, 55-58.
2. Jiang, H., Wang, Y., Huang, Y., Mulnix, A., Kadel, J., Cole, K. and Kanost, M.R. (1996) Organization of serpin gene-1 from Manduca sexta: evolution of a family of alternate exons encoding the reactive site loop. J. Biol. Chem. 271, 28017-28023.
3. Jiang, H. and Kanost, M.R. (1997) Characterization and functional analysis of twelve naturally occurring reactive site variants of serpin-1 from Manduca sexta. J. Biol. Chem. 272, 1082-1087.
4. Jiang, H., Wang, Y., Ma, C., and Kanost, M.R. (1997) Subunit composition of pro-phenol oxidase from Manduca sexta: molecular cloning of subunit proPO-p1. Insect Biochem. Mol. Biol. 27, 835-850.
5. Jiang, H., Wang, Y., Korochkina, S.E., Beneš, H., and Kanost, M.R. (1997) Molecular cloing of cDNAs for two pro-phenol oxidase subunits from the malaria vector, Anopheles gambiae. Insect Biochem. Mol. Biol. 27, 693-699.
6. Jiang, H., Wang, Y., and Kanost, M.R. (1998) Pro-phenol oxidase activating proteinase from an insect, Manduca sexta: a bacteria-inducible protein similar to Drosophila easter. Proc. Natl. Acad. Sci. USA. 95, 12220-12225.
7. Wang, Y., Jiang, H., and Kanost, M.R. (1999) Biological activity of Manduca sexta paralytic and plasmatocyte spreading peptide and primary structure of its hemolymph precursor. Insect Biochem. Mol. Biol. 29, 1075-1086.
8. Jiang, H. and Kanost, M.R. (2000) The clip-domain family of serine proteinases in arthropods. Insect Biochem. Mol. Biol. 30, 95-105.
9. Wang, Y., Jiang, H., and Kanost, M.R. (2001) Expression and purification of Manduca sexta prophenoloxidase-activating proteinase precursor (proPAP) from baculovirus-infected insect cells. Protein Express. Purif., 23, 328-337.
10. Yu, X-Q., Jiang, H., Wang, Y., and Kanost, M.R. (2003) Nonproteolytic serine proteinase homologs are involved in phenoloxidase activation in the tobacco hornworm, Manduca sexta. Insect Biochem. Mol. Biol. 33, 197-208
11. Ross, J., Jiang, H., Kanost, M.R., and Wang, Y. (2003) Serine proteases and their homologs in the Drosophila melanogaster genome: an initial analysis of sequence conservation and phylogenetic relationship. Gene, 304, 117-131.
12. Jiang, H., Wang, Y., Yu, X-Q., and Kanost, M.R. (2003) Prophenoloxidase-activating proteinase-2 (PAP-2) from hemolymph of Manduca sexta: a bacteria-inducible serine proteinase containing two clip domains. J. Biol. Chem. 278, 3552-3561.
13. Jiang, H., Wang, Y., Yu, X-Q., Zhu, Y., and Kanost, M.R. (2003) Prophenoloxidase-activating proteinase-3 (PAP-3) from Manduca sexta hemolymph: a clip-domain serine proteinase regulated by serpin-1J and serine proteinase homologs. Insect Biochem. Mol. Biol. 33, 1049-1060.
14. Zhu, Y., Wang, Y., Gorman, M., Jiang, H., and Kanost, M.R. (2003) Manduca sexta serpin-3 regulates prophenoloxidase activation in response to infection by inhibiting prophenoloxidase-activating proteinases. J. Biol. Chem. 47, 46556-46564.
15. Ji, C., Wang, Y., Ross, J., and Jiang, H. (2003) Expression and in vitro activation of Manduca sexta prophenoloxidase-activating proteinase-2 precursor (proPAP-2) from baculovirus-infected insect cells. Protein Express. Purif. 29, 235-243.
16. Jiang, H., Ma, C., Lu, Z., and Kanost, M.R. (2004) ß-1,3-glucan recognition protein-2 (ßGRP-2) from Manduca sexta: an acute-phase protein that binds ß-1,3-glucan and lipoteichoic acid to aggregate fungi and bacteria and stimulate prophenoloxidase activation. Insect Biochem. Mol. Biol. 34, 89-100.
17. Kanost, M.R., Jiang, H., and Yu, X. (2004) Innate immune responses of a lepidopteran insect, Manduca sexta. Immunol. Rev. 198, 97-105.
18. Wang, Y. and Jiang, H. (2004) Purification and characterization of Manduca sexta serpin-6: a serine proteinase inhibitor that selectively inhibits prophenoloxidase-activating proteinase-3. Insect Biochem. Mol. Biol. 34, 387-395.
19. Zhang, G., Lu, Z-Q, Jiang, H., and S. Asgari (2004) Negative regulation of prophenoloxidase (proPO) activation by a clip-domain serine proteinase homolog (SPH) from endoparasitoid venom. Insect Biochem. Mol. Biol. 34, 477-483.
20. Wang, Y. and Jiang, H. (2004) Prophenoloxidase (PPO) activation in Manduca sexta: an initial analysis of molecular interactions among PPO, PPO-activating proteinase-3 (PAP-3), and a cofactor. Insect Biochem. Mol. Biol. 34, 731-742.
21. Ji, C., Wang, Y., Guo, X., Hartson, S., and Jiang, H. (2004) A pattern recognition serine proteinase triggers the prophenoloxidase activation cascade in the tobacco hornworm,
Manduca sexta. J. Biol. Chem. 279, 34101-34106.
22. Gupta, S., Wang, Y., and Jiang, H. (2005) Purification and characterization of Manduca sexta prophenoloxidase-activating proteinase-1 (PAP-1), an enzyme involved in insect immune responses. Protein Exp. Purif. 39, 261-268.
23. Gupta, S., Wang, Y., and Jiang, H. (2005) Manduca sexta prophenoloxidase (proPO) activation requires proPO-activating proteinase (PAP) and serine proteinase homologs (SPHs) simultaneously. Insect Biochem. Mol. Biol., 35, 241-248.
24. Tong, Y., Jiang, H., and Kanost, M.R. (2005) Identification of plasma proteases inhibited by Manduca sexta serpin-4 and serpin-5 and their association with components of the prophenol oxidase activation pathway. J. Biol. Chem. 280, 14932-14942.
25. Zou, Z. and Jiang, H. (2005) Manduca sexta serpin-6 regulates immune serine proteinases PAP-3 and HP8: cDNA cloning, protein expression, inhibition kinetics, and function elucidation. J. Biol. Chem. 280, 14341-14348.
26. Zou, Z., Wang, Y., and Jiang, H. (2005) Manduca sexta prophenoloxidase activating proteinase-1 (PAP-1) gene: organization, expression, and regulation by immune and hormonal signals. Insect Biochem. Mol. Biol. 35, 627-636.
27. Jiang, H., Wang, Y., Gu, Y., Guo, X., Zou, Z., Scholz, F., Trenczek, T.E., and Kanost, M.R. (2005) Molecular identification of a bevy of serine proteinases in Manduca sexta hemolymph. Insect Biochem. Mol. Biol. 35, 931-943.
28. Zou, Z. and Jiang, H. (2005) Gene structure and transcriptional regulation of Manduca sexta prophenoloxidase-activating proteinase-3 (PAP-3), an immune protein containing two clip domains. Insect Mol. Biol. 14, 433-442.
29. Wang, Y., Zou, Z., and Jiang, H. (2006) An expansion of the dual clip-domain serine proteinase family in Manduca sexta: gene organization, expression, and evolution of prophenoloxidase-activating proteinase-2, hemolymph proteinase 12, and other related proteinases. Genomics. in press.
30. Wang, Y. and Jiang, H. (2006) Interaction of ß-1,3-glucan with its recognition protein activates hemolymph proteinase 14, an initiation enzyme of the prophenoloxidase activation system in Manduca sexta. J. Biol. Chem. manuscript submitted
