Summary
Several pathogenesis-related proteins, which are produced in plants submitted to stress, have been identified as chitinases. We have previously described that a potato basic chitinase strongly inhibited the activity of an aspartic protease isolated from the same source. In this work, we have tested the activity of two potato chitinases as protease inhibitors. A basic chitinase (ChiB) inhibited trypsin, chymotrypsin, subtilisin, proteinase K and a serine protease ofFusarium sp. An acidic one (ChiA) did not show inhibitory activity. The kinetics of trypsin inhibition by ChiB revealed different patterns of inhibition with azocasein and BAPNA. Metal ions affected differentially both activities, suggesting that ChiB is a bifunctional protein. These results and those reported by other authors suggest a new physiological role for pathogenesis-related proteins. However, results presented in this paper suggest that the protease inhibitor activity is not a general characteristic of potato chitinases.
Similar content being viewed by others
References
Abeles, F.B., R.P. Bosshart, L.E. Forrence & W.H. Habig. 1971. Preparation and purification of glucanase and chitinase from bean leaves.Plant Physiology 47: 129–134.
Andreu, A., C. Tonón, M. van Damme, M. Huarte & G. Daleo, 1998. Effect of glucans from different races ofPhytophthora infestans on defense reactions in potato tuber.European Journal of Plant Pathology 104: 777–783.
Adams, D.J., B.E. Causier, K.J. Mellor, V. Keer, R. Milling & J. Dada, 1993. Regulation of chitin synthase and chitinase in fungi. In: R.A.A. Muzzarelli (Ed.), Chitin enzymology. European Chitin Society, Lyon, France and Ancona, Italy, pp. 15–26.
Geoffroy, P., M. Legrand & B. Fritig, 1990. Isolation and characterization of a proteinaceous inhibitor of microbial proteases induced during the hypersensitive reaction of tobacco to tobacco mosaic virus.Molecular Plant-Microbe Interactions 3: 327–333.
Green, T.R. & C.A. Ryan, 1972. Wound-induced proteinase inhibitor in plant leaves: a possible defense mechanism against insects.Science 175: 776–777.
Guevara, M.G., C.R. Oliva, M. Machinandiarena & G.R. Daleo, 1999. Purification and properties of an aspartic protease from potato tuber that is inhibited by a basic chitinase.Physiologia Plantarum 106: 164–169.
Heitz, T., P. Geoffroy, B. Fritig & M. Legrand, 1999. The PR-6 family: proteinase inhibitors in plant-microbe and plant-insect interactions. In: S.K. Datta & S. Muthukrishnan (Eds), Pathogenesis-related proteins in plants. CRC Press Inc., Boca Raton, FL, pp. 131–155.
Kassell, B., 1970. Inhibitors of Proteolytic Enzymes. In: G.E. Perlmann & L. Lorand (Eds), Proteolytic Enzymes. Methods in Enzymology, London, New York, Vol XIX, pp. 839–905.
Laemmli, U.K., 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4.Nature 227: 680–685.
Lorito, M., R.M. Broadway, C.K. Hayes, S.L. Woo, C. Noviello, D.L. Williams & G.E. Harmann, 1994. Proteinase inhibitors from plants as a novel class of fungicides.Molecular Plant-Microbe Interactions 7: 525–527.
Mauch, F., B. Mauch-Mani & T. Boller, 1988. Antifungal hydrolases in pea tissue. II. Inhibition of fungal growth by combination of chitinase and β-1,3 glucanase.Plant Physiology 88: 936–942.
Oakley, B.R., D.R. Kirsch & N.R. Morris, 1980. A simplified ultra sensitive silver stain for detecting proteins in polyacrilamide gels.Analytical Biochemistry 105: 361–363.
Oliva, C.R., 1993. Cambios metabólicos en discos de papa frente a diferentes situaciones de estrés. PhD Thesis, Universidad Nacional de Mar del Plata.
Reissig, J., J. Strominger & L.F. Leloir, 1955. A modified colorimetric method for the estimation of N-acetylaminosugars.Journal of Biological Chemistry 217: 959–966.
Richardson, M., 1977. The proteinase inhibitors of plants and microorganisms.Phytochemistry 16: 159–169.
Rickauer, M., J. Fournier & M.T. Esquerre-Tugaye, 1989. Induction of proteinase inhibitors in tobacco cell suspension culture by elicitors ofPhytophthora parasitica cv. nicotianae.Plant Physiology 90: 1065–1070.
Smith, P.K., R.I. Krohn, G.T. Hermanson, A.K. Maallia, F.H. Gartner, M.D. Provenzano, E.K. Fujimoto, N.M. Goeke, B.J. Olson & D.C. Klenk, 1985. Measurement of protein using bicinchoninic acid.Analytical Biochemistry 150: 76–85.
Valueva, T.A., T.A. Revina, G.V. Kladnitskaya & V.V. Mosolv, 1998. Kunitz-type proteinase inhibitors from intact andPhytophthora-infected potato tubers.FEBS Letters 426: 131–134.
Wessels, J. & J. Sietsma, 1981. Fungal cell walls: a survey. In: W. Tanner & F. Loewus (Eds), Plant Carbohydrates II. Encyclopedia of Plant Physiology, New Series, Springer-Verlag, Berlin, Vol. 13 B, pp. 352–394.
Yamakawa, T., S. Miyata, N. Ogawa, N. Koshikawa, H. Yasumitsu, T. Kanamori & K. Miyazaki, 1998. cDNA cloning of the novel trypsin inhibitor with similarity to pathogenesis-related proteins, and its frequent expression in human brain cancer cells.Biochimica et Biophysica Acta 1395: 202–208.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Machinandiarena, M., Castillo, M., Olivieri, F. et al. Protease inhibitor activity is associated to a basic chitinase from potato but not to an acidic one. Potato Res 44, 187–195 (2001). https://doi.org/10.1007/BF02410105
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF02410105