Abstract
The sequence of a trypsin inhibitor, isolated from wheat endosperm, is reported. The primary structure was obtained by automatic sequence analysis of the S-alkylated protein and of purified peptides derived from chemical cleavage by cyanogen bromide and digestion withStaphylococcus aureus V8 protease. This protein, named wheat trypsin inhibitor (WTI), which is comprised of a total of 71 amino acid residues, has 12 cysteines, all involved in disulfide bridges. The primary site of interaction (reactive site) with bovine trypsin has been identified as the dipeptide arginyl-methionyl at positions 19 and 20. WTI has a high degree of sequence identity with a number of serine proteinase inhibitors isolated from both cereal and leguminous plants. On the basis of the findings presented, this protein has been classified as a single-headed trypsin inhibitor of Bowman-Birk type.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Ary, M. B., Shewry, P. R., and Richardson, M. (1988).FEBS Lett. 229, 111–118.
Birk, Y., Gertler, A., and Khalef, S. (1963).Biochem. J. 87, 281–284.
Bode, W., and Huber, R. (1992).Eur. J. Biochem. 204, 433–451.
Boisen, S., and Djurtoft, R. (1981).Cereal Chem. 58, 460–463.
Boisen, S. (1983).Acta Agric. Scand. 33, 369–381.
Bowman, D. E. (1946).Proc. Soc. Exp. Biol. Med. 63, 547–550.
Brown, W. E., Takio, K., Titani, K., and Ryan, C. A. (1985).Biochemistry 24, 2105–2108.
Buonocore, V., and Silano, V. (1986). InNutritional and Toxicological Significance of Enzyme Inhibitors in Foods (Friedman, M., ed.), Plenum Press, New York, pp. 483–507.
Carrano, L., Nitti, G., Buonocore, V., Caporale, C., and Poerio, E. (1989).Plant Sci. 65, 25–31.
Ellman, G. L. (1959).Arch. Biochem. Biophys. 82, 70–77.
Eckelkamp, C., Ehmann, B., and Schopfer, P. (1993).FEBS Lett. 323, 73–76.
Garcia-Olmedo, F., Salcedo, G., Sanchez-Monge, R., Gomez, L., Royo, J., and Carbonero, P. (1987).Oxf. Suru. Plant Mol. Cell Biol. 4, 275–334.
Gross, E., and Witkop, B. (1961).J. Am. Chem. Soc. 83, 1510–1511.
Hilder, V. A., Barker, R. F., Samour, R. A., Gatehouse, A. M. R., Gatehouse, J. A., and Boulter, D. (1969).Plant Mol. Biol. 13, 701–710.
Ikenaka, T., and Norioka, S. (1986). InProteinase Inhibitors (Barrett, A. J., and Salvesen, G., eds.), Elsevier Science Publishers, Amsterdam, pp. 361–374.
Ishikawa, C., Nakamura, S., Watanabe, K., and Takahashi, K. (1979).FEBS Lett. 99, 97–100.
Joubert, F. J., Kruger, H., Townshend, G. S., and Botes, D. P. (1979).Eur. J. Biochem. 97, 85–91.
Joubert, F. J. (1982).S. Afr. J. Chem. 35, 72–76.
Jubert, F. J. (1984).Phytochemistry 23, 957–961.
Kiyohara, T., Yokota, K., Masaki, Y., Matsui, O., Iwasaki, T., and Yoshikawa, M. (1981).J. Biochem. 90, 721–728.
Laemmli, U. K. (1970).Nature 227, 680–685.
Laskowski, M., Jr., and Kato, I. (1980).Annu. Rev. Biochem. 49, 593–626.
Laskowsi, M., Jr. (1986). InNutritional and Toxicological Significance of Enzyme Inhibitors in Foods (Friedman, M., ed.), Plenum Press, New York, pp. 1–17.
Mikes, O., Holeysovsky, V., Tomasek, V., and Sorm, F. (1966).Biochem. Biophys. Res. Commun. 24, 346–352.
Mitsunaga, T. (1979).J. Nutr. Sci. Vitaminol. 25, 43–52.
Mitsunaga, T., Kimura, Y., and Shimizu, M. (1982).J. Nutr. Sci. Vitaminol. 28, 419–429.
Nagasue, A., Fukamachi, H., Ikenaga, H., and Funatsu, G. (1988).Agric. Biol. Chem. 52, 1505–1514.
Norioka, S., and Ikenaka, T. (1983).J. Biochem. 94, 589–599.
Odani, S., and Ikenaka, T. (1972).J. Biochem. 71, 839–848.
Odani, S., and Ikenaka, T. (1977).J. Biochem. 82, 1523–1531.
Odani, S., and Ikenaka, T. (1978).J. Biochem. 83, 737–745.
Odani, S., Koide, T., and Ono, T. (1986).J. Biochem. 100, 975–983.
Poerio, E., Carrano, L., Garzillo, A. M., and Buonocore, V. (1989).Phytochemistry 28, 1307–1311.
Reisfieid, R. A., Lewis, U. J., and Williams, D. E. (1962).Nature 195, 281–283.
Richardson, M. (1991). InMethods of Plant Biochemistry (Rogers, L. J., ed.), Vol. 5, Academic Press, London, pp. 259–305.
Ryan, C. A. (1990).Annu. Rev. Phytopathol. 28, 425–449.
Shimokawa, Y., Kuromizu, K., Araki, T., Ohata, J., and Abe, O. (1984).Eur. J. Biochem. 143, 677–684.
Stevens, F. C., Wuery, S., and Krahu, J. (1974). InProteinase Inhibitors, Bayer Symp. V (Fritz, H., Tschesche, H., Greene, L. J., and Truscheit, E., eds.), Springer Verlag, Berlin, pp. 344–354.
Tashiro, M., Hashino, K., Shiozaki, M., Ibuki, F., and Maki, Z. (1987).J. Biochem. 102, 297–306.
Tashiro, M., Asao, T., Hirata, C., Takahashi, K., and Kanamori, M. (1990).J. Biochem. 108, 669–672.
Wilson, K. A., and Laskowski, M., Sr. (1975).J. Biol. Chem. 250, 4261–4267.
Zhang, Y. S., Los, S., Tan, F. L., Chi, C. W., Xu, L. X., and Zhang, A. L. (1982).Sci. Sinica 25, 268–277.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Poerio, E., Caporale, C., Carrano, L. et al. The amino acid sequence and reactive site of a single-headed trypsin inhibitor from wheat endosperm. J Protein Chem 13, 187–194 (1994). https://doi.org/10.1007/BF01891977
Received:
Published:
Issue Date:
DOI: https://doi.org/10.1007/BF01891977