Isolation of cell-surface glycopeptides from bovine cerebral cortex that inhibit cell growth and protein synthesis in normal but not in transformed cells

Abstract

Glycopeptides were isolated from the cell surfaces of bovine cerebral cortex that could inhibit increase in cell numbers in tissue culture and cellular protein synthesis. This cell growth inhibition apparently affected all cells exposed, could completely block cell division in a reversible manner and synchronized BHK-21 cell cultures. Polyoma-virus-transformed BHK-21 cells were completely insensitive to the inhibitor. Fractionation of the inhibitor on a Bio-Gel P-100 column revealed two peaks of biologically active material eluting at apparent molecular weights of 45 000 and 10 000 with A 1cm,280 1% 11.0. Affinity purification of the inhibitory fractions on a Ulex europaeus agglutination I lectin column resulted in retention of the inhibitory activity, suggesting the inhibitor material was a glycopeptide. Subsequent elution with 0.10 M-fucose resulted in a 244-fold increase in the specific biological activity over the starting material. Although purified from bovine brain, the material could inhibit baby-hamster kidney cell protein synthesis by 50% at a concentration of 5×10(6) molecules per target cell. Analysis by competitive radioimmunoassay or immunoadsorption indicated that the bovine inhibitor was structurally related to, although not necessarily identical with, a similar inhibitory glycopeptide preparation that we had previously isolated from mouse brain.