Journal of Dairy Research

Original Articles

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

Esben S. Sørensena1 and Torben E. Petersena1

a1 Protein Chemistry Laboratory, Department of Molecular Biology, University of Aarhus, The Science Park, Gustav Wieds Vej 10, DK-8000 Aarhus C, Denmark

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.

(Received July 02 1992)

(Accepted October 12 1992)