Isolation of a clone of Chinese hamster ovary cells deficient in plant lectin-binding sites

Abstract

Ricin, a galactose-binding lectin with potent cytotoxic activity, was used to select a clone of Chinese hamster ovary cells with altered plant lectin-binding properties. The clone (15B) is 80-fold less sensitive to the toxic action of ricin than the parent line. In the absence of ricin, it grows both in monolayer and suspension culture with a normal generation time. Plating efficiency, however, is significantly reduced. Relative to the parent cells, its binding of the Ricinus communis lectins, Phaseolus vulgaris erythroagglutinating phytohemagglutinin, Abrus precatorius phytohemagglutinin, and soybean phytohemagglutinin is less than 7%, while binding of lentil phytohemagglutinin, wheat-germ agglutinin, and mushroom phytohemagglutinin is 17%, 40%, and 109%, respectively. In contrast, its concanavalin A binding is increased by 70%. Consistent with these alterations, crude membrane preparations of the 15B cells were found to contain the same sugars as the parent-cell membranes but in different proportions. The 15B membranes have 28% less sialic acid, 38% less N-acetylglucosamine, 49% less galactose, the same amount of N-acetylgalactosamine, and 53% more mannose than the membranes of the parent cells.