Oklahoma State University

Dr. Haobo Jiang, Professor



B.S., East China University of Chem Tech, 1985
Ph.D., Kansas State University, 1994
haobo.jiang@okstate.edu


Links:

Manduca sexta EST contigs of hemolymph proteins involved in immune responses

Serine Protease (SP) and Serine Protease Homologous (SPH) Proteins in Drosophila melanogaster

A multiple sequence alignment of the glucanase-like proteins in invertebrates

Sequence alignment of the catalytic domains of Manduca sexta HPs and other arthropod serine proteinases with known functions

Teaching:

ENTO/PLP 5623     Advanced Biotechnology Methods


Lab members, students, and collaborators:

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

Picheng Zhao, Graduate Research Assistant
Native of Shanghai, China
2001  B.S., East China University of Science and Technology, Biochemical engineering

Fan Yang, Graduate Research Assistant
Native of Jiangsu, China
2004  B.S., East China University of Science and Technology, Food Science and Technology
2007  M.S., East China University of Science and Technology, Microbial and Biochemical Pharmacology

Niranji Sumathipala, Graduate Research Assistant
Native of Sri Lanka
2005  B.S., University of Colombo, Biochemistry and Molecular Biology

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:  (All 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., 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.

    6. Jiang, H. and Kanost, M.R. (2000) The clip-domain family of serine proteinases in arthropods. Insect Biochem. Mol. Biol. 30, 95-105.

    7. 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

    8. 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.

    9. 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.

    10. 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.

    11. 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.

    12. 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.

    13. 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.

    14. 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.

    15. 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.

    16. 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.

    17. 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.

    18. 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. 87, 399-409.

    19. 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. 281, 9271-9278.

    20. Wang, Y., Chen, T., Rayaprolu, S., Zou, Z., Xia, Q., Xiang, Z., and Jiang, H. (2007) Proteolytic activation of pro-spätzle is required for the induced transcription of antimicrobial peptide genes in lepidopteran insects. Dev. Com. Immunol. 31, 1002-1012.

    21. Wang, Y. and Jiang, H. (2007) Reconstitution of a branch of Manduca sexta prophenoloxi- dase activation cascade in vitro: Snake-like hemolymph proteinase 21 cleaved by HP14 activates prophenoloxidase-activating proteinase-2 precursor. Insect Biochem. Mol. Biol. 37, 1015-1025.

    22. Gorman, M.J., Wang, Y., Jiang, H., and Kanost, M.R. (2007) Manduca sexta hemolymph proteinase 21 activates prophenoloxidase activating proteinase 3 in an insect innate immune response proteinase cascade. J. Biol. Chem. 282, 11742-11749.

    23. Waterhouse, R.M., Kriventseva, E.V., Meister, S., Xi, Z., Alvarez, K.S., Bartholomay, L.C., Barillas-Mury, C., Bian, G., Blandin, S., Christensen, B.M., Dong, Y., Jiang, H., Kanost, M.R., Koutsos, A.C., Levashina, E.A., Li, J., Ligoxygakis, P., Maccallum, R.M., Mayhew, G.F., Mendes, A., Michel, K., Osta, M.A., Paskewitz, S., Shin, S.W., Vlachou, D., Wang, L., Wei, W., Zheng, L., Zou, Z., Severson, D.W., Raikhel, A.S., Kafatos, F.C., Dimopoulos, G., Zdobnov, E.M., Christophides, G.K. (2007) Evolutionary dynamics of immune-related genes and pathways in disease-vector mosquitoes. Science. 316 (5832), 1738-1743.

    24. Zhao, P., Li, J., Wang, Y., and Jiang, H. (2007) Broad-spectrum antimicrobial activity of the reactive compounds generated in vitro by Manduca sexta phenoloxidase. Insect Biochem. Mol. Biol. 37, 952-959.

    25. Huang, R., Lu, Z., Dai, H., Vander Velde, D., Prakash, O., and Jiang, H. (2007) The solution structure of the clip domains from Manduca sexta prophenoloxidase activating proteinase-2. Biochemistry 46, 11431-11439.

    26. Zou, Z., Evans, J., Lu, Z., Zhao, P., Williams, M., Sumathipala, N., Hetru, C., Hultmark, D., and Jiang, H. (2007) Comparative genome analysis of the Tribolium immune system. Genome Biol. 8, R177.

    27. Jiang, H. (2008) The biochemical basis of antimicrobial responses in Manduca sexta. Insect Science 15, 53-66.

    28. Zou, Z., Najar, F., Wang, Y., Roe, B., and Jiang, H. (2008) Analysis of Manduca sexta expressed sequence tags by pyrosequencing. Insect Biochem. Mol. Biol. 38, 677-682.

    29. Wang, Y. and Jiang, H. (2008) A positive feedback mechanism in the Manduca sexta prophenoloxidase activation. Insect Biochem. Mol. Biol. 38, 763-769.

    30. Lu, Z, Beck, M.H., Wang, Y., Jiang, H., and Strand, M.R. (2008) The viral protein EGF1.0 is a dual activity inhibitor of prophenoloxidase activating proteinases 1 and 3 from Manduca sexta. J. Biol. Chem. 283, 21325-21333.



    • Division of Agricultural Sciences and Natural Resources