Isolation of Two New Proteins from Bovine Colostrum Which Stimulate Epidermal Growth Factor-Dependent Colony Formation of NRK-49F Cells

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Abstract
Defatted bovine colostrum and decaseinated whey contain biological activity similar to that of TGF-β in that they stimulate the anchorage-independent growth of NRK-49F cells in the presence of EGF. Two new proteins with such activity, termed BC-1 and BC-2, were isolated from decaseinated colostrum by a sequence of DEAE-Sephacel chromatography, Sephadex G-100 gel filtration in 1 M acetic acid, Sephadex G-50 re-gelfiltration in 8 M urea-1 M acetic acid and reverse-phase FPLCs. The two proteins showed distinctly different molecular weight from that of TGF-β1. BC-1 was a small Mr peptide of 8.5 kD. BC-2 had molecular mass of 46 kD, which yielded peptides of 46, 40, and 6 kD on reduction. In contrast to TGF-β, both proteins did not lose their biological activity on reduction. Both BC-1 and BC-2 suppress DNA synthesis in concanavalin A-stimulated thymocytes. These results suggest that BC-1 and BC-2 belong to a new class of mitogen/inhibitors, though their biological activities resemble those of TGF-β.