Abstract
Trypsin inhibitors of two varieties of Bauhinia variegata seeds have been isolated and characterized. Bauhinia variegata candida trypsin inhibitor (BvcTI) and B. variegata lilac trypsin inhibitor (BvlTI) are proteins with Mr of about 20,000 without free sulfhydryl groups. Amino acid analysis shows a high content of aspartic acid, glutamic acid, serine, and glycine, and a low content of histidine, tyrosine, methionine, and lysine in both inhibitors. Isoelectric focusing for both varieties detected three isoforms (pI 4.85, 5.00, and 5.15), which were resolved by HPLC procedure. The trypsin inhibitors show Ki values of 6.9 and 1.2 nM for BvcTI and BvlTI, respectively. The N-terminal sequences of the three trypsin inhibitor isoforms from both varieties of Bauhinia variegata and the complete amino acid sequence of B. variegata var. candida L. trypsin inhibitor isoform 3 (BvcTI-3) are presented. The sequences have been determined by automated Edman degradation of the reduced and carboxymethylated proteins of the peptides resulting from Staphylococcus aureus protease and trypsin digestion. BvcTI-3 is composed of 167 residues and has a calculated molecular mass of 18,529. Homology studies with other trypsin inhibitors show that BvcTI-3 belongs to the Kunitz family. The putative active site encompasses Arg (63)–Ile (64).
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
REFERENCES
Edman, P., and Begg, G. (1967). Eur. J. Biochem. 1, 80–91.
Heinrikson, R. L., and Meridith, S. C. (1984). Anal. Biochem. 136, 65.
Kim, S.-H., Hara, S., Hase, S., Ikenaka, T., Toda, H., Kitamura, K., and Kaizuma, N. (1985). J. Biochem. 98, 435–448.
Knight, C.-G. (1986). In Protease Inhibitors (Barret, A. J., and Salvesen, G., eds), Elsevier, Amsterdam, pp. 23–51.
Laemmli, U. K. (1970). Nature 227, 680.
Marangoni, S., Ghiso, J., Sampaio, S. V., Arantes, E. C., Giglio, J. R., Oliveira, B., and Frangione, B. J. (1990). J. Protein Chem. 9, 595–601.
Marangoni, S., Toyama, M. H., Arantes, E. C., Giglio, J. R., Silva, C. A., Carneiro, E. C., Gonçalves, A. A., and Oliveira, B. (1995). Biochim. Biophys. Acta 1243, 309–314.
Oliva, M. L. V., Sampaio, M. U., and Sampaio, C. A. M. (1987). Braz. J. Med. Biol. Res. 20, 767.
Richardson, M. (1977). Phytochemistry 16, 159.
Richardson, M. (1991). Meth. Plant Biochem. 5, 259.
Ryan, C. A. (1973). Annu. Rev. Plant Physiol. 24, 173–196.
Sampaio, C. A. M., Oliva, M. L. V., Sampaio, M. U., Batista, I. F. C., Buena, N. R., Tanaka, A. S., Auerswald, E. A., and Fritz, H. (1996). Immunopharmacology, 32, 62–66.
Sampaio, C. A. M., Sampaio, M. U., and Prado, E. S. (1984). Hoppe-Seyler's Z. Physiol. Chem. 365, 297.
Weder, J. K. P. (1991). In Proceedings 1st Brazilian Congress on Proteins, (Oliveira, B., and Sgarbieri, V., eds.), Editora Unicamp, Campinas, Brazil, pp. 29–55.
Wu, H.-C., and Lin, J.-Y. (1993). J. Biochem. 113, 258–263.
Xavier-Filho, J., and Campos, F. A. P. (1989). In Protease Inhibitors and Toxicants of Plant Origin (Cheek, P. R., ed.), CRC Press. Boca Raton, Florida, Vol. 3, pp. 1–27.
Yamamoto, M., Hara, S., and Ikenaka, T. (1983). J. Biochem. 94, 849–863.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Ciero, L.D., Oliva, M.L.V., Torquato, R. et al. The Complete Amino Acid Sequence of a Trypsin Inhibitor from Bauhinia variegata var. candida Seeds. J Protein Chem 17, 827–834 (1998). https://doi.org/10.1023/A:1020734519908
Published:
Issue Date:
DOI: https://doi.org/10.1023/A:1020734519908