Hostname: page-component-cc8bf7c57-ksm4s Total loading time: 0 Render date: 2024-12-11T23:04:00.245Z Has data issue: false hasContentIssue false
  • English
  • Français

Purification and characterization of three proteins isolated from the proteose peptone fraction of bovine milk

Published online by Cambridge University Press:  01 June 2009

Esben S. Sørensen  and
Torben E. Petersen
Esben S. Sørensen
Affiliation:
Protein Chemistry Laboratory, Department of Molecular Biology, University of Aarhus, The Science Park, Gustav Wieds Vej 10, DK-8000 Aarhus C, Denmark
Torben E. Petersen
Affiliation:
Protein Chemistry Laboratory, Department of Molecular Biology, University of Aarhus, The Science Park, Gustav Wieds Vej 10, DK-8000 Aarhus C, Denmark
Get access Rights & Permissions [Opens in a new window]

Summary

Three major proteins from the proteose peptone of bovine milk were purified by Sephadex G-75 gel chromatography, Q-Sepharose ion-exchange and additional Sephadex G-75 gel chromatography in the presence of urea. From their mobility in a gradient SDS-PAGE, the proteins were found to have molecular masses of 17, 28 and 60 kDa. The N-terminal amino acid sequence of the 17 kDa protein was found to be homologous with a camel whey protein. This protein has not previously been described in bovine milk. From the SDS-PAGE results, the 28 kDa protein was judged to be the major protein of proteose peptone, contributing ~ 25% of the total. The N-terminal amino acid sequence showed no homology to any known protein sequence, but the amino acid composition indicated that the 28 kDa protein is identical with the PP3 component from the proteose peptone fraction of bovine milk, or part of it. The 60 kDa protein was found to be bovine osteopontin, a very highly phosphorylated protein with an Arg-Gly-Asp sequence which mediates cell attachment.

Type
Original Articles
Information
Journal of Dairy Research , Volume 60 , Issue 2 , May 1993 , pp. 189 - 197
Copyright
Copyright © Proprietors of Journal of Dairy Research 1993

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

REFERENCES

Andrews, A. T. 1978 a The composition, structure and origin of proteose-peptone component 5 of bovine milk. European Journal of Biochemistry 90 59–65Google Scholar
Andrews, A. T. 1978 b The composition, structure and origin of proteose-peptone component 8F of bovine milk. European Journal of Biochemistry 90 67–71Google Scholar
Andrews, A. T. & Alichanidis, E. 1983 Proteolysis of caseins and the proteose-peptone fraction of bovine milk. Journal of Dairy Research 50 275–290Google Scholar
Barkholt, V. & Jensen, A. L. 1989 Amino acid analysis: determination of cysteine plus half-cystine in proteins after hydrochloric acid hydrolysis with a disulfide compound as additive. Analytical Biochemistry 177 318–322Google Scholar
Beg, O. U., Von Bahr-Lindström, H., Zaidi, Z. H. & Jörnvall, H. 1987 Characterization of a heterogeneous camel milk whey non-casein protein. FEBS Letters 216 270–274Google Scholar
Butler, W T. 1989 The nature and significance of osteopontin. Connective Tissue Research 23 123–136Google Scholar
Craig, A. M., Bowden, G. T., Chambers, A. F., Spearman, M. A., Greenberg, A. H., Wright, J. A., McLeod, M. & Denhardt, D. T. 1990 Secreted phosphoprotein mRNA is induced during multi-stage carcinogenesis in mouse skin and correlates with the metastatic potential of murine fibroblasts. International Journal of Cancer 46 133–137CrossRefGoogle ScholarPubMed
Craig, A. M. & Denhardt, D. T. 1991 The murine gene encoding secreted phosphoprotein 1 (osteopontin): promoter structure, activity, and induction in vivo by estrogen and progesterone. Gene 100 163–171Google Scholar
Craig, A. M., Smith, J. H., & Denhardt, D. T. 1989 Osteopontin, a transformation-associated cell adhesion phosphoprotein, is induced by 12-O-tetradecanoylphorbol-13-acetate in mouse epidermis. Journal of Biological Chemistry 264 9682–9689Google Scholar
Eigel, W. N. & Keenan, T. W. 1979 Identification of proteose-peptone component 8-slow as a plasmin-derived fragment of bovine β-casein. International Journal of Biochemistry 10 529–535CrossRefGoogle ScholarPubMed
Fet, V., Dickinson, M. E. & Hogan, B. L. M. 1989 Localization of the mouse gene for secreted phosphoprotein 1 (Spp-1) (2ar, osteopontin, bone sialoprotein I. 44-k Dal bone phosphoprotein, tumor-secreted phosphoprotein) to cliromosome 5, closely linked to Ric (Rickettsia resistance) Genomics 5 375–377Google Scholar
Franzén, A. & Heinegård, D. 1985 Isolation and characterization of two sialoproteins present only in bone calcified matrix. Biochemical Journal 232 715–724CrossRefGoogle ScholarPubMed
Jack, L. J. W. & Mather, I. H. 1990 Cloning and analysis of cDNA encoding bovine butyrophilin, an apical glycoprotein expressed in mammary tissue and secreted in association with the milk-fat globule membrane during lactation. Journal of Biological Chemistry 265 14481–14486Google Scholar
Kanno, C. 1989 a Purification and separation of multiple forms of lactophorin from bovine milk whey and their immunological and electrophoretic properties. Journal of Dairy Science 72 883–891CrossRefGoogle ScholarPubMed
Kanno, C. 1989 b Characterization of multiple forms of lactophorin isolated from bovine milk whey. Journal of Dairy Science 72 1732–1739Google Scholar
Kerr, J. M., Fisher, L. W., Termine, J. D. & Young, M. F. 1991 The cDNA cloning and RNA distribution of bovine ostcopontin. Gene 108 237–243Google Scholar
Kester, J. J. & Brunner, J. R. 1982 Milk fat-globule membrane as possible origin of proteose-peptone glycoproteins. Journal of Dairy Science 65 2241–2252Google Scholar
Kolar, C. W. & Brunner, J. R. 1970 Proteose-peptone fraction of bovine milk: lacteal serum components 5 and 8 casein-associated glycoproteins. Journal of Dairy Science 53 997–1008CrossRefGoogle ScholarPubMed
Laemmli, U. K. 1970 Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227 680–685Google Scholar
Larson, B. L. & Rolleri, G. D. 1955 Heat denaturation of specific serum proteins in milk. Journal of Dairy Science 38 351–360CrossRefGoogle Scholar
Mark, M. P., Prince, C. W., Gay, S., Austin, R. L., Bhown, M., Finkelman, R. D. & Butler, W. T. 1987 a A comparative immunoeytochcmical study on the subcellular distribution of a 44 kDal bone phosphoprotein and bone γ-carboxyglutamic acid (Gla)-containing protein in osteoblasts. Journal of Hone and Mineral Research 2 337–346Google Scholar
Mark, M. P., Prince, C. W., Gay, S., Austin, R. L. & Butler, W. T. 1988 44-kDal bone phosphoprotein (osteopontin) antigenicity at ectopic sites in newborn rats: kidney and nervous tissues. Cell and Tissue Research 251 23–30Google Scholar
Mark, M. P., Prince, C. W., Oosawa, T., Gay, S., Bronckers, A. L. J. J. & Butler, W. T. 1987 b Immunohistoehemical demonstration of a 44 kDal phosphoprotein in developing rat bones. Journal of Histochemistry and Cytochemistry 35 707–715Google Scholar
Nejjar, Y., Pâquet, D., Godbillon, G. & Le Deaut, J. Y. 1986 Immunological relationship between the hydrophobic fraction of proteose-peptone and the milk fat globule membrane of bovine milk. International Journal of Biochemistry 18 893–900CrossRefGoogle ScholarPubMed
Ng, W. C., Brunner, J. R. & Rhee, K. C. 1970 Proteose-peptone fraction of bovine milk: lacteum scrum component 3—a whey glycoprotein. Journal of Dairy Science 53 987–996Google Scholar
Nomura, S., Wills, A. J., Edwards, D. R., Heath, J. K. & Hogan, B. L. M. 1988 Developmental expression of 2ar (osteopontin) and SPARC (osteonectin) RNA as revealed by in situ hybridization. Journal of Cell Biology 106 441–450Google Scholar
Oldberg, Å, Franzén, A. & Heinegård, D. 1986 Cloning and sequence analysis of rat bone sialoprotein (ostcopontin) cDNA reveals an Arg-Gly-Asp cell-binding sequence. Proceedings of the National Academy of Sciences of the USA 83 8819–8823Google Scholar
Påquet, D. 1989 [Review: the proteose peptone fraction of milk.] Lait 69 1–21Google Scholar
Påquet, D., Nejjar, Y. & Linden, G. 1988 Study of a hydrophobic protein fraction isolated from milk proteose-peptone. Journal of Dairy Science 71 1464–1471Google Scholar
Rowlands, S. J. 1937 The soluble protein fraction of milk. Journal of Dairy Research 8 6–14Google Scholar
Rowlands, S. J. 1938 The precipitation of the proteins in milk. 1. Casein. 2. Total proteins. 3. Globulins, 4. Albumin and proteose-peptone. Journal of Dairy Research 9 30–41CrossRefGoogle Scholar
Senger, D. R., Perruzzi, C. A., Papadopoulos, A. & Tenen, D. G. 1989 Purification of a human milk protein closely similar to tumor-secreted phosphoprotcins and osteopontin. Biochimica et Biophysica. Acta 996 43–48Google Scholar
Shiraga, H., Min, W., VanDusen, W. J., Clayman, M. D., Miner, D., Terrell, C. H., Sherbotie, J. R., Foreman, J. W., Przysiecki, C., Neilson, E. G. & Hoyer, J. R. 1992 Inhibition of calcium oxalate crystal growth in nitro by uropontin: another member of the aspartic acid-rich protein superfamily. Proceedings of the National Academy of Sciences of the USA 89 426–430Google Scholar
Yoon, K., Buenaga, R. & Rodan, G. A. 1987 Tissue specificity and developmental expression of rat osteopontin. Biochemical and Biophysical Research Communications 148 1129–1136Google Scholar
Young, M. F., Kerr, J. M., Termine, J. D., Wewer, U. M., Wang, M. G., McBride, O. W. & Fisher, L. W. 1990 cDNA cloning, mRNA distribution and heterogeneity, chromosomal location, and RFLP analysis of human ostcopontin (OPN). Genomics 7 491–502Google Scholar