Skip to main content

Salivary Protease Inhibitors with Non Anti-Hemostatic Functions

  • Chapter
  • First Online:
Toxins and Hemostasis

Abstract

Arthropod saliva regulates proteolysis in the sites of bite(s). Secreted salivary inhibitors target different proteolytic enzymes (proteases) that have a pivotal role in vertebrate hemostasis, immunity and inflammation. The aim of this chapter is to give an overview of the salivary protease inhibitors that affect the latter two physiological procedures. Functional studies, mainly in ticks, have demonstrated many potent arthropod salivary inhibitors of cysteine proteases (cathepsins) and serine proteases (elastase and tryptase). Emphasis is given to the function of these inhibitors and more specifically to the mechanism by which they facilitate hematophagy.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Chapter
USD 29.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 169.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 219.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 219.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  • Azzolini, S.S., Santos, J.M., Souza, A.F., Torquato, R.J., Hirata, I.Y., Andreotti, R., Tanaka, A.S., 2004. Purification, characterization, and cloning of a serine proteinase inhibitor from the ectoparasite Haematobia irritans irritans (Diptera: Muscidae). Exp. Parasitol. 106, 103–109.

    Article  PubMed  CAS  Google Scholar 

  • Belaaouaj, A., McCarthy, R., Baumann, M., Gao, Z., Ley, T.J., Abraham, S.N., Shapiro, S.D., 1998. Mice lacking neutrophil elastase reveal impaired host defense against gram negative bacterial sepsis. Nat. Med. 4, 615–618.

    Article  PubMed  CAS  Google Scholar 

  • Blair, R.J., Meng, H., Marchese, M.J., Ren, S., Schwartz, L.B., Tonnesen, M.G., Gruber, B.L., 1997. Human mast cells stimulate vascular tube formation. Tryptase is a novel, potent angiogenic factor. J. Clin. Invest. 99, 2691–2700.

    Article  PubMed  CAS  Google Scholar 

  • Corral-Rodriguez, M.A., Macedo-Ribeiro, S., Barbosa Pereira, P.J., Fuentes-Prior, P., 2009. Tick-derived Kunitz-type inhibitors as antihemostatic factors. Insect Biochem. Mol. Biol. 39, 579–595.

    Article  PubMed  CAS  Google Scholar 

  • Corvera, C.U., Dery, O., McConalogue, K., Bohm, S.K., Khitin, L.M., Caughey, G.H., Payan, D.G., Bunnett, N.W., 1997. Mast cell tryptase regulates rat colonic myocytes through proteinase-activated receptor 2. J. Clin. Invest. 100, 1383–1393.

    Article  PubMed  CAS  Google Scholar 

  • Coussens, L.M., Raymond, W.W., Bergers, G., Laig-Webster, M., Behrendtsen, O., Werb, Z., Caughey, G.H., Hanahan, D., 1999. Inflammatory mast cells up-regulate angiogenesis during squamous epithelial carcinogenesis. Genes Dev. 13, 1382–1397.

    Article  PubMed  CAS  Google Scholar 

  • Dainichi, T., Maekawa, Y., Ishii, K., Zhang, T., Nashed, B.F., Sakai, T., Takashima, M., Himeno, K., 2001. Nippocystatin, a cysteine protease inhibitor from Nippostrongylus brasiliensis, inhibits antigen processing and modulates antigen-specific immune response. Infect. Immun. 69, 7380–7386.

    Article  PubMed  CAS  Google Scholar 

  • Felbor, U., Dreier, L., Bryant, R.A., Ploegh, H.L., Olsen, B.R., Mothes, W., 2000. Secreted cathepsin L generates endostatin from collagen XVIII. EMBO J. 19, 1187–1194.

    Article  PubMed  CAS  Google Scholar 

  • Francischetti, I.M., Mather, T.N., Ribeiro, J.M., 2005. Tick saliva is a potent inhibitor of endothelial cell proliferation and angiogenesis. Thromb. Haemost. 94, 167–174.

    PubMed  CAS  Google Scholar 

  • Francischetti, I.M., Sá-Nunes, A., Mans, B.J., Santos, I.M., Ribeiro, J.M., 2009. The role of saliva in tick feeding. Front. Biosci. 14, 2051–2088.

    Article  PubMed  CAS  Google Scholar 

  • Gadher, S.J., Eyre, D.R., Duance, V.C., Wotton, S.F., Heck, L.W., Schmid, T.M., Woolley, D.E., 1988. Susceptibility of cartilage collagens type II, IX, X, and XI to human synovial collagenase and neutrophil elastase. Eur. J. Biochem. 175, 1–7.

    Article  PubMed  CAS  Google Scholar 

  • Ganz, T., Metcalf, J.A., Gallin, J.I., Boxer, L.A., Lehrer, R.I., 1988. Microbicidal/cytotoxic proteins of neutrophils are deficient in two disorders: Chediak-Higashi syndrome and “specific” granule deficiency. J. Clin. Invest. 82, 552–556.

    Article  PubMed  CAS  Google Scholar 

  • Goto, S.G., Denlinger, D.L., 2002. Genes encoding two cystatins in the flesh fly Sarcophaga crassipalpis and their distinct expression patterns in relation to pupal diapause. Gene 292, 121–127.

    Article  PubMed  CAS  Google Scholar 

  • Gruber, B.L., Marchese, M.J., Suzuki, K., Schwartz, L.B., Okada, Y., Nagase, H., Ramamurthy, N.S., 1989. Synovial procollagenase activation by human mast cell tryptase dependence upon matrix metalloproteinase 3 activation. J. Clin. Invest. 84, 1657–1662.

    Article  PubMed  CAS  Google Scholar 

  • Grunclova, L., Horn, M., Vancova, M., Sojka, D., Franta, Z., Mares, M., Kopacek, P., 2006. Two secreted cystatins of the soft tick Ornithodoros moubata: differential expression pattern and inhibitory specificity. Biol. Chem. 387, 1635–1644.

    Article  PubMed  CAS  Google Scholar 

  • He, S., Aslam, A., Gaca, M.D., He, Y., Buckley, M.G., Hollenberg, M.D., Walls, A.F., 2004. Inhibitors of tryptase as mast cell-stabilizing agents in the human airways: effects of tryptase and other agonists of proteinase-activated receptor 2 on histamine release. J. Pharmacol. Exp. Ther. 309, 119–126.

    Article  PubMed  CAS  Google Scholar 

  • Heck, L.W., Blackburn, W.D., Irwin, M.H., Abrahamson, D.R., 1990. Degradation of basement membrane laminin by human neutrophil elastase and cathepsin G. Am. J. Pathol. 136, 1267–1274.

    PubMed  CAS  Google Scholar 

  • Honey, K., Rudensky, A.Y., 2003. Lysosomal cysteine proteases regulate antigen presentation. Nat. Rev. Immunol. 3, 472–482.

    Article  PubMed  CAS  Google Scholar 

  • Huang, C., De Sanctis, G.T., O’Brien, P.J., Mizgerd, J.P., Friend, D.S., Drazen, J.M., Brass, L.F., Stevens, R.L., 2001. Evaluation of the substrate specificity of human mast cell tryptase beta I and demonstration of its importance in bacterial infections of the lung. J. Biol. Chem. 276, 26276–26284.

    Article  PubMed  CAS  Google Scholar 

  • Ide, H., Itoh, H., Yoshida, E., Kobayashi, T., Tomita, M., Maruyama, H., Osada, Y., Nakahata, T., Nawa, Y., 1999. Immunohistochemical demonstration of inter-alpha-trypsin inhibitor light chain (bikunin) in human mast cells. Cell Tissue Res. 297, 149–154.

    Article  PubMed  CAS  Google Scholar 

  • Kaminska, R., Helisalmi, P., Harvima, R.J., Naukkarinen, A., Horsmanheimo, M., Harvima, I.T., 1999. Focal dermal-epidermal separation and fibronectin cleavage in basement membrane by human mast cell tryptase. J. Invest. Dermatol. 113, 567–573.

    Article  PubMed  CAS  Google Scholar 

  • Karim, S., Miller, N.J., Valenzuela, J., Sauer, J.R., Mather, T.N., 2005. RNAi-mediated gene silencing to assess the role of synaptobrevin and cystatin in tick blood feeding. Biochem. Biophys. Res. Commun. 334, 1336–1342.

    Article  PubMed  CAS  Google Scholar 

  • Kido, H., Fukusen, N., Katunuma, N., 1988. Antibodies and inhibitor of chymase are incorporated into mast cell granules and inhibit histamine release. Biol. Chem. Hoppe-Seyler. 369(Suppl), 95–100.

    PubMed  CAS  Google Scholar 

  • Koh, C.Y., Kini, R.M., 2009. Molecular diversity of anticoagulants from haematophagous animals. Thromb. Haemost. 102, 437–453.

    PubMed  CAS  Google Scholar 

  • Kotsyfakis, M., Anderson, J.M., Andersen, J.F., Calvo, E., Francischetti, I.M., Mather, T.N., Valenzuela, J.G., Ribeiro, J.M., 2008. Cutting edge: immunity against a "silent" salivary antigen of the Lyme vector Ixodes scapularis impairs its ability to feed. J. Immunol. 181, 5209–5212.

    PubMed  CAS  Google Scholar 

  • Kotsyfakis, M., Karim, S., Andersen, J.F., Mather, T.N., Ribeiro, J.M., 2007. Selective cysteine protease inhibition contributes to blood-feeding success of the tick Ixodes scapularis. J. Biol. Chem. 282, 29256–29263.

    Article  PubMed  CAS  Google Scholar 

  • Kotsyfakis, M., Sá-Nunes, A., Francischetti, I.M., Mather, T.N., Andersen, J.F., Ribeiro, J.M., 2006. Antiinflammatory and immunosuppressive activity of sialostatin L, a salivary cystatin from the tick Ixodes scapularis. J. Biol. Chem. 281, 26298–26307.

    Article  PubMed  CAS  Google Scholar 

  • Kozik, A., Moore, R.B., Potempa, J., Imamura, T., Rapala-Kozik, M., Travis, J., 1998. A novel mechanism for bradykinin production at inflammatory sites. Diverse effects of a mixture of neutrophil elastase and mast cell tryptase versus tissue and plasma kallikreins on native and oxidized kininogens. J. Biol. Chem. 273, 33224–33229.

    Article  PubMed  CAS  Google Scholar 

  • Leboulle, G., Crippa, M., Decrem, Y., Mejri, N., Brossard, M., Bollen, A., Godfroid, E., 2002a. Characterization of a novel salivary immunosuppressive protein from Ixodes ricinus ticks. J. Biol. Chem. 277, 10083–10089.

    Article  PubMed  CAS  Google Scholar 

  • Leboulle, G., Rochez, C., Louahed, J., Ruti, B., Brossard, M., Bollen, A., Godfroid, E., 2002b. Isolation of Ixodes ricinus salivary gland mRNA encoding factors induced during blood feeding. Am. J. Trop. Med. Hyg. 66, 225–233.

    PubMed  CAS  Google Scholar 

  • Lima, C.A., Sasaki, S.D., Tanaka, A.S., 2006. Bmcystatin, a cysteine proteinase inhibitor characterized from the tick Boophilus microplus. Biochem. Biophys. Res. Commun. 347, 44–50.

    Article  PubMed  CAS  Google Scholar 

  • Lombardi, G., Burzyn, D., Mundinano, J., Berguer, P., Bekinschtein, P., Costa, H., Castillo, L.F., Goldman, A., Meiss, R., Piazzon, I., Nepomnaschy, I., 2005. Cathepsin-L influences the expression of extracellular matrix in lymphoid organs and plays a role in the regulation of thymic output and of peripheral T cell number. J. Immunol. 174, 7022–7032.

    PubMed  CAS  Google Scholar 

  • Molinari, J.F., Moore, W.R., Clark, J., Tanaka, R., Butterfield, J.H., Abraham, W.M., 1995. Role of tryptase in immediate cutaneous responses in allergic sheep. J. Appl. Physiol. 79, 1966–1970.

    PubMed  CAS  Google Scholar 

  • Molinari, J.F., Scuri, M., Moore, W.R., Clark, J., Tanaka, R., Abraham, W.M., 1996. Inhaled tryptase causes bronchoconstriction in sheep via histamine release. Am. J. Respir. Crit. Care Med. 154, 649–653.

    Article  PubMed  CAS  Google Scholar 

  • Paesen, G.C., Siebold, C., Harlos, K., Peacey, M.F., Nuttall, P.A., Stuart, D.I., 2007. A tick protein with a modified Kunitz fold inhibits human tryptase. J. Mol. Biol. 368, 1172–1186.

    Article  PubMed  CAS  Google Scholar 

  • Prevot, P.P., Adam, B., Boudjeltia, K.Z., Brossard, M., Lins, L., Cauchie, P., Brasseur, R., Vanhaeverbeek, M., Vanhamme, L., Godfroid, E., 2006. Anti-hemostatic effects of a serpin from the saliva of the tick Ixodes ricinus. J. Biol. Chem. 281, 26361–26369.

    Article  PubMed  CAS  Google Scholar 

  • Prevot, P.P., Beschin, A., Lins, L., Beaufays, J., Grosjean, A., Bruys, L., Adam, B., Brossard, M., Brasseur, R., Zouaoui Boudjeltia, K., Vanhamme, L., Godfroid, E., 2009. Exosites mediate the anti-inflammatory effects of a multifunctional serpin from the saliva of the tick Ixodes ricinus. FEBS J. 276, 3235–3246.

    Article  PubMed  CAS  Google Scholar 

  • Prevot, P.P., Couvreur, B., Denis, V., Brossard, M., Vanhamme, L., Godfroid, E., 2007. Protective immunity against Ixodes ricinus induced by a salivary serpin. Vaccine 25, 3284–3292.

    Article  PubMed  CAS  Google Scholar 

  • Reddy, V.Y., Zhang, Q.Y., Weiss, S.J., 1995. Pericellular mobilization of the tissue-destructive cysteine proteinases, cathepsins B, L, and S, by human monocyte-derived macrophages. Proc. Natl. Acad. Sci. U.S.A. 92, 3849–3853.

    Article  PubMed  CAS  Google Scholar 

  • Reinheckel, T., Hagemann, S., Dollwet-Mack, S., Martinez, E., Lohmuller, T., Zlatkovic, G., Tobin, D.J., Maas-Szabowski, N., Peters, C., 2005. The lysosomal cysteine protease cathepsin L regulates keratinocyte proliferation by control of growth factor recycling. J. Cell Sci. 118, 3387–3395.

    Article  PubMed  CAS  Google Scholar 

  • Ribeiro, J.M., Arca, B., Lombardo, F., Calvo, E., Phan, V.M., Chandra, P.K., Wikel, S.K., 2007. An annotated catalogue of salivary gland transcripts in the adult female mosquito, Aedes aegypti. BMC Genomics 8, 6.

    Article  PubMed  Google Scholar 

  • Ribeiro, J.M., Francischetti, I.M., 2003. Role of arthropod saliva in blood feeding: sialome and post-sialome perspectives. Annu. Rev. Entomol. 48, 73–88.

    Article  PubMed  CAS  Google Scholar 

  • Rice, A., Banda, M.J., 1995. Neutrophil elastase processing of gelatinase A is mediated by extracellular matrix. Biochemistry 34, 9249–9256.

    Article  PubMed  CAS  Google Scholar 

  • Sá-Nunes, A., Bafica, A., Antonelli, L.R., Choi, E.Y., Francischetti, I.M., Andersen, J.F., Shi, G.P., Chavakis, T., Ribeiro, J.M., Kotsyfakis, M., 2009. The immunomodulatory action of sialostatin L on dendritic cells reveals its potential to interfere with autoimmunity. J. Immunol. 182, 7422–7429.

    Article  PubMed  Google Scholar 

  • Sakai, K., Long, S.D., Pettit, D.A., Cabral, G.A., Schwartz, L.B., 1996. Expression and purification of recombinant human tryptase in a baculovirus system. Protein Expr. Purif. 7, 67–73.

    Article  PubMed  CAS  Google Scholar 

  • Sant’Anna Azzolini, S., Sasaki, S.D., Torquato, R.J., Andreotti, R., Andreotti, E., Tanaka, A.S., 2003. Rhipicephalus sanguineus trypsin inhibitors present in the tick larvae: isolation, characterization, and partial primary structure determination. Arch. Biochem. Biophys. 417, 176–182.

    Article  PubMed  Google Scholar 

  • Sasaki, S.D., Azzolini, S.S., Hirata, I.Y., Andreotti, R., Tanaka, A.S., 2004. Boophilus microplus tick larvae, a rich source of Kunitz type serine proteinase inhibitors. Biochimie 86:643–649.

    Article  PubMed  CAS  Google Scholar 

  • Sasaki, S.D., de Lima, C.A., Lovato, D.V., Juliano, M.A., Torquato, R.J., Tanaka, A.S., 2008. BmSI-7, a novel subtilisin inhibitor from Boophilus microplus, with activity toward Pr1 proteases from the fungus Metarhizium anisopliae. Exp. Parasitol. 118, 214–220.

    Article  PubMed  CAS  Google Scholar 

  • Serveau-Avesque, C., Martino, M.F., Herve-Grepinet, V., Hazouard, E., Gauthier, F., Diot, E., Lalmanach, G., 2006. Active cathepsins B, H, K, L and S in human inflammatory bronchoalveolar lavage fluids. Biol. Cell 98, 15–22.

    Article  PubMed  CAS  Google Scholar 

  • Shamamian, P., Schwartz, J.D., Pocock, B.J., Monea, S., Whiting, D., Marcus, S.G., Mignatti, P., 2001. Activation of progelatinase A (MMP-2) by neutrophil elastase, cathepsin G, and proteinase-3: a role for inflammatory cells in tumor invasion and angiogenesis. J. Cell. Physiol. 189, 197–206.

    Article  PubMed  CAS  Google Scholar 

  • Stack, M.S., Johnson, D.A., 1994. Human mast cell tryptase activates single-chain urinary-type plasminogen activator (pro-urokinase). J. Biol. Chem. 269, 9416–9419.

    PubMed  CAS  Google Scholar 

  • Tanaka, A.S., Andreotti, R., Gomes, A., Torquato, R.J., Sampaio, M.U., Sampaio, C.A., 1999. A double headed serine proteinase inhibitor--human plasma kallikrein and elastase inhibitor—from Boophilus microplus larvae. Immunopharmacology 45, 171–177.

    Article  PubMed  CAS  Google Scholar 

  • Turk, B., Turk, D., Turk, V., 2000. Lysosomal cysteine proteases: more than scavengers. Biochim. Biophys. Acta 1477, 98–111.

    Article  PubMed  CAS  Google Scholar 

  • Yamaji, K., Tsuji, N., Miyoshi, T., Islam, M.K., Hatta, T., Alim, M.A., Anisuzzaman, M., Kushibiki, S., Fujisaki, K., 2009. A salivary cystatin, HlSC-1, from the ixodid tick Haemaphysalis longicornis play roles in the blood-feeding processes. Parasitol. Res. 106(1), 61–68.

    Google Scholar 

  • Zavasnik-Bergant, T., Turk, B., 2007. Cysteine proteases: destruction ability versus immunomodulation capacity in immune cells. Biol. Chem. 388, 1141–1149.

    Article  PubMed  CAS  Google Scholar 

  • Zhou, J., Liao, M., Ueda, M., Gong, H., Xuan, X., Fujisaki, K., 2009. Characterization of an intracellular cystatin homolog from the tick Haemaphysalis longicornis. Vet. Parasitol. 160, 180–183.

    Article  PubMed  CAS  Google Scholar 

  • Zhou, J., Ueda, M., Umemiya, R., Battsetseg, B., Boldbaatar, D., Xuan, X., Fujisaki, K., 2006. A secreted cystatin from the tick Haemaphysalis longicornis and its distinct expression patterns in relation to innate immunity. Insect Biochem. Mol. Biol. 36, 527–535.

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgements

This work was supported by the Intramural Research Program of the Division of Intramural Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, by the grant No. IAA600960811 from the Grant Agency of the Academy of Sciences of the Czech Republic and by the Research Center No. LC06009 from the Ministry of Education, Youth and Sports of the Czech Republic. We thank Professor R. Manjunatha Kini for the fruitful discussions and comments on the manuscript, Dr. Koh Cho Yeow for preparing the figure and NIAID intramural editor Brenda Rae Marshall for assistance.

Because I.M.B.F. is a government employee and this is a government work, the work is in the public domain in the United States. Notwithstanding any other agreements, the NIH reserves the right to provide the work to PubMedCentral for display and use by the public, and PubMedCentral may tag or modify the work consistent with its customary practices. You can establish rights outside of the U.S. subject to a government use license.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Michalis Kotsyfakis .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2010 Springer Science+Business Media B.V.

About this chapter

Cite this chapter

Chmelař, J., Francischetti, I.M., Kotsyfakis, M. (2010). Salivary Protease Inhibitors with Non Anti-Hemostatic Functions. In: Kini, R., Clemetson, K., Markland, F., McLane, M., Morita, T. (eds) Toxins and Hemostasis. Springer, Dordrecht. https://doi.org/10.1007/978-90-481-9295-3_10

Download citation

Publish with us

Policies and ethics