Abstract
Bovine serum albumin has been covalently labeled with Remazol brilliant blue R to provide a substrate for a convenient spectrophotometric assay for protein precipitants. The blue protein is especially useful for measuring protein precipitation by vegetable tannins because its absorption maximum is at a wavelength where plant pigments exhibit minimum absorption. Blue BSA has been used to determine, by competition experiments, the relative affinity of various proteins for tannins. A procedure for purifying condensed tannin from commercially available quebracho extract is described.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Armstrong, G.S. 1983. A study of tannin-protein interactions. PhD thesis, Purdue University, West Lafayette, Indiana.
Asquith, T.N., Izuno, C.C., andButler, L.G. 1983. Characterization of the condensed tannin (proanthocyanidin) from a group II sorghum.J. Agric. Food Chem. 31:1299–1303.
Bate-Smith, E.C. 1973. Haemanalysis of tannins: The concept of relative astringency. Phytochemistry 12:907–912.
Becker, P., andMartin, J.S. 1982. Protein-precipitating capacity of tannins inShorea (Diptero-carpaceae) seedling leaves.J. Chem. Ecol. 8:1353–1367.
BioRad Bulletin No. 1089. 1982. The BioRad Silver Stain.
Bradford, M.M. 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.Anal. Biochem. 72:248–254.
Bullard, R.W., York, J.O., andKilburn, S.R. 1981. Polyphenolic changes in ripening bird-resistant sorghums.J. Agric. Food Chem. 29:973–984.
Butler, L.G., Price, M.L., andBrotherton, J.E. 1982. Vanillin assay for proanthocyanidins (condensed tannins): Modification of the solvent for estimation of the degree of polymerization.J. Agric. Food Chem. 30:1087–1089.
Butler, L.G., Riedl, D.J., Lebryk, D.G., andBlytt, H.J. 1984. Interaction of proteins with sorghum tannin: Mechanism, specificity and significance.J. Am. Oil Chem. Soc. 61:916–920.
Calderon, P., Van Buren, J., andRobinson, W.B. 1968. Factors influencing the formation of precipitates and hazes by gelatin and condensed and hydrolyzable tannins.J. Agric. Food Chem. 16:479–483.
Daiber, K.H. 1975. Enzyme inhibition by polyphenols of sorghum grain and malt.J. Sci. Food Agric. 26:1399–1411.
Davis, A.B., andHoseney, R.C. 1979. Grain sorghum condensed tannins. II. Preharvest changes.Cereal Chem. 56:310–316.
Daytner, A., andFinnimore, E.D. 1973. A prestaining method for the quantitative assay of proteins on polyacrylamide gels.Anal. Biochem. 55:479–491.
Earp, C.G., Akinobala, J.O., Ring, S.H., andRooney, L.W. 1981. Evaluation of several methods to determine tannins in sorghums with varying kernel characteristics.Cereal Chem. 58:234–238.
Gray, M. 1978. Absorption of polyphenols by polyvinylpyrrolidone and polystyrene resins.Phytochemistry 17:495–497.
Gupta, R.K., andHaslam, E. 1980. Pages 15–24,in Proceedings of the Symposium on “Polyphenols in Cereals and Legumes,” 36th Annual Institute of Food Technologists Meeting, St. Louis, Missouri, June 10–13, 1979; International Research Center; Ottawa, Ontario, IDRC-14-e.
Hagerman, A.E., andButler, L.G. 1978. Protein precipitation method for the quantitative determination of tannins.J. Agric. Food Chem. 26:809–812.
Hagerman, A.E., andButler, L.G. 1980a. Determination of protein in tannin-protein precipitates.J. Agric. Food Chem. 28:944–947.
Hagerman, A.E., andButler, L.G. 1980b. Condensed tannin purification and characterization of tannin-associated proteins.J. Agric. Food Chem. 28:947–952.
Hagerman, A.E., andButler, L.G. 1981. The specificity of proanthocyanidin-protein interactions.J. Biol. Chem. 256:4494–4497.
Laemeli, U.K. 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4.Nature 227:680–685.
Lowry, O.H., Rosebrough, N.J., Farr, A.L., andRandall, R.J. 1951. Protein measurement with the Folin-phenol reagent.J. Biol. Chem. 193:265–275.
Martin, J.S., andMartin, M.M. 1983. Tannin assays in ecological studies. Precipitation of ribulose-1-5-biphosphate carboxylase/oxygenase by tannic acid, quebracho and oak foliage extracts.J. Chem. Ecol. 9:285–294.
McManus, J.P., Davis, K.G., Lilley, T.H., andHaslam, E. 1981. The association of proteins with polyphenols.J. Chem. Soc. Chem. Commun. Perkin Trans. 00:309–311.
Mehansho, H., andCarlson, D.M. 1983. Induction of protein and glycoprotein synthesis in rat submandibular gland by isoproterenol.J Biol. Chem. 258:6616–6620
Mehansho, H., Hagerman, A.E., Clements, S., Butler, L.G., Rogler, J., andCarlson, D.M. 1983. Modulation of proline-rich protein biosynthesis in rat parotid glands by sorghums.Proc. Natl. Acad. Sci. U.S.A. 80:3948–3952.
Mehansho, H., Rogler, J., Butler, L.G., andCarlson, D.M. 1985. An unusual growth inhibiting effect of tannins on hamsters.Fed. Proc., 44:1860.
Porter, L.J., andWoodruff, J. 1984. Haemanalysis: The relative astringency of proanthocyanidin polymers.Phytochemistry 23:1255–1256.
Rinderknecht, H., Geokas, M.C., Silverman, P., andHaverback, B.J. 1968. A new ultrasensitive method for the determination of proteolytic activity.Clin. Chim. Acta 21:197–203.
Roux, D.G. 1957. Some recent advances in the identification of leucoanthocyanins and the chemistry of condensed tannins.Nature 180:973–975.
Schultz, J.C., Baldwin, I.T., andNorthnagle, P.J. 1981. Hemoglobin as a binding substrate in the quantitative analysis of plant tannins.J. Agric. Food Chem. 29:823–826.
Strumayer, D.H., andMalin, M.J. 1975. Condensed tannins in grain sorghum: Isolation, fractionation, and characterization.J. Agric. Food Chem. 23:909–914.
Wilson, M.F. 1984. Comparison of tannin levels in developing fruit buds of two orchard pear varieties using two techniques, Folin-Denis and protein precipitation assays.J. Chem. Ecol. 10:493–498.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Asquith, T.N., Butler, L.G. Use of dye-labeled protein as spectrophotometric assay for protein precipitants such as tannin. J Chem Ecol 11, 1535–1544 (1985). https://doi.org/10.1007/BF01012199
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF01012199