Hostname: page-component-78c5997874-m6dg7 Total loading time: 0 Render date: 2024-11-19T16:53:42.578Z Has data issue: false hasContentIssue false
  • English
  • Français

Purification of goats' milk casein by reversed-phase high-performance liquid chromatography and identification of αs1-casein

Published online by Cambridge University Press:  01 June 2009

Jan Mikkelsen ,
Peter Højrup  and
Jens Knudsen
Jan Mikkelsen
Affiliation:
Institute of Biochemistry, DK-5230 Odense M, Denmark
Peter Højrup
Affiliation:
Institute of Molecular Biology, University of Odense, DK-5230 Odense M, Denmark
Jens Knudsen
Affiliation:
Institute of Biochemistry, DK-5230 Odense M, Denmark
Get access Rights & Permissions [Opens in a new window]

Summary

Goats’ milk caseins were separated into four components in a single run using reversed-phase gradient high-performance liquid chromatography. The purity of the isolated components was checked by sodium dodecyl sulphate polyacrylamide gel electrophoresis, amino acid analysis and determination of the N-terminal residue. By a comparison with previously published results for goats’ milk caseins the four peaks were identified as κ-, αs1-, αs2- and β-casein. In order to confirm the existence of αsl-casein in goats’ milk, this component was sequenced for 44 steps, revealing a sequence homologous to bovine αsl-casein and almost identical to the N-terminal sequence previously published by Boulanger et al. (1984).

Type
Original Articles
Information
Journal of Dairy Research , Volume 54 , Issue 3 , August 1987 , pp. 361 - 367
Copyright
Copyright © Proprietors of Journal of Dairy Research 1987

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., Taylor, M. D. & Owen, A. J. 1985 Rapid analysis of bovine milk proteins by fast protein liquid chromatography. Journal of Chromatography 348 177–185CrossRefGoogle ScholarPubMed
Boulanger, A., Grosclaude, F. & Mahé, M. -F. 1984 [Polymorphism of αsl- and αs2-caseins in the goat (Capra hircus).] Génétique, Sélection, Evolution 16 157–175CrossRefGoogle Scholar
Brignon, G., Chtourou, A. & Ribadeau-Dumas, B. 1985 Preparation and amino acid sequence of human κ-casein. FEBS Letters 188 48–54CrossRefGoogle ScholarPubMed
Davies, D. T. & Law, A. J. R. 1977 An improved method for the quantitative fractionation of casein mixtures using ion-exchange chromatography. Journal of Dairy Research 44 213–221CrossRefGoogle Scholar
Esch, F. S. 1984 Polypeptide microsequence analysis with the commercially available gas phase sequencer. Analytical Biochemistry 136 39–47CrossRefGoogle ScholarPubMed
Gray, W. 1967 Sequential degradation plus dansylation. Methods in Enzymology 11 469–475CrossRefGoogle Scholar
Hunkapiller, M. W. & Hood, L. E. 1983 Analysis of phenylthiohydantoins by ultrasensitive gradient high-performance liquid chromatography. Methods in Enzymology 91 486–493CrossRefGoogle ScholarPubMed
Laemmli, U. K. 1970 Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227 680–685CrossRefGoogle ScholarPubMed
Lowry, O. H., Rosebrough, N. J., Farr, A. L. & Randall, R. J. 1951 Protein measurement with the folin phenol reagent. Journal of Biological Chemistry 193 265–275CrossRefGoogle ScholarPubMed
Mercier, J. C., Grosclaude, F. & Ribadeau-Dumas, B. 1971 [Structure of bovine αsl-casein.] European Journal of Biochemistry 23 41–51CrossRefGoogle Scholar
Picon, P. 1985 The application of high-performance ion-exchange chromatography for the analysis of bovine milk proteins. Experientia 41 958–960Google Scholar
Richardson, B. C. & Creamer, L. K. 1974 Comparative micelle structure. III. The isolation and chemical characterization of caprine β1-casein and β2-casein. Biochimica et Biophysica Acta 365 133–137CrossRefGoogle Scholar
Richardson, B. C., Creamer, L. K. & Munford, R. E. 1973 Comparative micelle structure. I. The isolation and chemical characterization of caprine κ-casein. Biochimica et Biophysica Acta 310 111–117CrossRefGoogle ScholarPubMed
Schacterle, G. R. & Pollack, R. L. 1973 A simplified method for the quantitative assay of small amounts of protein in biological material. Analytical Biochemistry 51 654–655CrossRefGoogle Scholar
Thompson, M. P. 1966 DEAE-cellulose-urea chromatography of casein in the presence of 2-mercapto-ethanol. Journal of Dairy Science 49 792–795CrossRefGoogle Scholar