Abstract
Samples of homozygous bovine serum transferrins have been prepared and their purity has been ascertained by immunological techniques and electrophoretic analysis in SDS. Measurements of carbohydrate composition show that no significant differences exist among the phenotypic variants AA, D1D1, D2D2, and EE. Chromatography of transferrin AA on DEAE-cellulose separated four subfractions, each of which corresponded well with one band obtained by polyacrylamide gel electrophoresis. Carbohydrate analyses of the individual subfractions did not show significant differences in sialic acid, hexose, or hexosamine contents. After desialylation with neuraminidase, each subfraction was converted to a major band and a minor band on gel electrophoresis. From the relative band positions of the desialylated transferrins, it was concluded that possession of sialyl residues by bovine transferrin is not the primary cause of electrophoretic multiplicity. Rather, sialic acid masks an underlying heterogeneity which most likely resides within the polypeptide chain. Further characterization of this heterogeneity will best be undertaken with the isolated asialotransferrin subfractions.
Similar content being viewed by others
References
Aminoff, D. (1961). Methods for the quantitative estimation of N-acetylneuraminic acid and their application to hydrolysates of sialomucoids. Biochem. J. 81384.
Ashton, G. C. (1958). Genetics of beta-globulin polymorphism in British cattle. Nature 182370.
Ashton, G. C., Gilmour, D. G., Kiddy, C. A., and Kristjansson, F. K. (1967). Proposals on nomenclature of protein polymorphisms in farm livestock. Genetics 56353.
Ashwell, G., and Morell, A. G. (1974). The role of surface carbohydrates in the hepatic recognition and transport of circulating glycoproteins. Adv. Enzymol. 4199.
Cassidy, J. T., Jourdian, G. W., and Roseman, S. (1965). The Sialic Acids. VI: Purification and properties of sialidase from Clostridium perfringens. J. Biol. Chem. 2403501.
Chen, S.-H., and Sutton, H. E. (1967). Bovine transferrins: Sialic acid and the complex phenotype. Genetics 56425.
Clarke, J. T. (1964). Simplified “disc” (polyacrylamide gel) electrophoresis. Ann. N.Y. Acad. Sci. 121428.
Dubois, M., Gilles, K. A., Hamilton, J. K., Rebers, P. A., and Smith, F. A. (1956). Colorimetric method for determination of sugars and related substances. Anal. Chem. 28350.
Dunker, A. K., and Rueckert, R. R. (1969). Observations on molecular weight determinations on polyacrylamide gel. J. Biol. Chem. 2445074.
Efremov, G. D., Smith, L. L., Barton, B. P., and Huisman, T. H. J. (1971). Studies on bovine transferrin; isolation and partial characterisation. Anim. Blood Grps. Biochem. Genet. 2159.
Graham, I., and Williams, J. (1975). A comparison of glycopeptides from the transferrins of several species. Biochem. J. 145263.
Hatton, M. W. C., and Regoeczi, E. (1973). A simple method for the purification of commercial neuraminidase preparations free from proteases. Biochim. Biophys. Acta 327114.
Hatton, M. W. C., Regoeczi, E., and Wong, K.-L. (1974). Studies of the metabolism of asialotransferrins: Relationship between the carbohydrate composition of bovine, canine and porcine asialotransferrins and their metabolic behaviour in the rabbit. Can. J. Biochem. 52845.
Hudson, B. G., Ohno, M., Brockway, W. J., and Castellino, F. J. (1973). Chemical and physical properties of serum transferrins from several species. Biochemistry 121047.
Jamieson, A. (1965). The genetics of transferrins in cattle. Heredity 20419.
Kristjansson, F. K., and Hickman, C. G. (1965). Subdivision of the allele TfD for transferrins in Holstein and Ayrshire cattle. Genetics 52627.
McFarlane, A. S. (1958). Efficient trace-labelling of protein with iodine. Nature 1853.
Mohr, E. and Schramm, G. (1960). Reinigung und Charakterisierung der Neuraminidase aus Vibrio cholerae. Z. Naturforsch. 156568.
Ouchterlony, O. (1948). Antigen-antibody reaction in gels. Ark. Kemi Mineral. Geol. 26(141.
Parker, W. C., and Bearn, A. G. (1962). Studies on the transferrins of adult serum, cord serum and cerebrospinal fluid. J. Exp. Med. 11583.
Regoeczi, E. (1975). Hepatic uptake of asialoglycoproteins in vivo: Quantification using a dual-isotope technique. J. Nucl. Biol. Med. 19149.
Regoeczi, E., Hatton, M. W. C., and Wong, K.-L. (1974). Studies of the metabolism of asialotransferrins: Potentiation of the catabolism of human asialotransferrin in the rabbit. Can. J. Biochem. 52155.
Regoeczi, E., Wong, K.-L., and Hatton, M. W. C. (1975). Studies of the metabolism of asialotransferrins: The metabolic heterogeneity of human asialotransferrin. Can. J. Biochem. 531070.
Richardson, N. E., Buttress, N., Feinstein, A., Stratil, A., and Spooner, R. L. (1973). Structural studies on individual components of bovine transferrin. Biochem. J. 13587.
Rondle, C. M. J., and Morgan, W. T. J. (1955). The determination of glucosamine and galactosamine. Biochem. J. 61586.
Scharmann, W., Brückler, J., and Blobel, H. (1971). Wirkung bakterieller Neuraminidasen auf Transferrin vom Menschen, Rind und Kaninchen. Biochim. Biophys. Acta 229136.
Scheidegger, J. J. (1955). Une micro-méthode de l'immunoélectrophorése. Int. Arch. Allergy Appl. Immunol. 7103.
Schultze, H. E., and Heremans, J. F. (1966). Molecular Biology of Human Proteins, Vol. 1, Elsevier, Amsterdam, p. 216.
Shapiro, A. L., Vinuela, E., and Maizel, J. V. (1967). Molecular weight estimation of polypeptide chains by electrophoresis in SDS-polyacrylamide gels. Biochem. Biophys. Res. Commun. 28815.
Smithies, O., and Hickman, C. G. (1958). Inherited variations in the serum proteins of cattle. Genetics 43374.
Stratil, A., and Spooner, R. L. (1971). Isolation and properties of individual components of cattle transferrin: The role of sialic acid. Biochem. Genet. 5347.
Wong, K.-L., Charlwood, P. A., Hatton, M. W. C., and Regoeczi, E. (1974). Studies of the metabolism of asialotransferrins: Evidence that transferrin does not undergo desialylation in vivo. Clin. Sci. Mol. Med. 46763.
Author information
Authors and Affiliations
Additional information
This research was supported by Grants MT-4074 and MA-5554 from the Medical Research Council of Canada and a Senior Fellowship (to M. W. C. H.) from the Ontario Heart Foundation.
Rights and permissions
About this article
Cite this article
Hatton, M.W.C., Regoeczi, E., Wong, K.L. et al. Bovine serum transferrin phenotypes AA, D1D1, D2D2, EE: Their carbohydrate compositions and electrophoretic multiplicity. Biochem Genet 15, 621–640 (1977). https://doi.org/10.1007/BF00484094
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF00484094