Two variants of B16 mouse
melanoma cells, selected for their resistance to toxic levels of
wheat germ agglutinin isolectin 1 (WGA-1) in serum-free medium, showed by chromosome analysis that they are still mouse cell-lines, continue to produce
melanin, and are less tumorigenic in mice than the parent B16 cells. The variants showed a marked decrease in cell agglutination with the
wheat germ lectin and a slight increase in cell agglutination with
concanavalin A. The binding of 125I-labeled
wheat germ agglutinin to the two variant lines was likewise decreased over a 10(3)-fold range of
lectin concentrations. Terminal sialyl residues were critical in WGA-1 binding to the wild-type cells. The binding data indicated a decrease in high-affinity binding as well as a decrease in the total number of binding sites in the variants.
Polyacrylamide gel electrophoresis, followed by affinity staining with 125I-wheat germ
agglutinin, showed alterations in the
wheat germ agglutinin-binding
glycoproteins in the variants compared to those of the parent cell line. However,
lactoperoxidase-catalyzed iodination revealed a similar
cell-surface protein pattern among the three cell lines. Radioactive
glycoproteins secreted or shed by the three cell lines grown in the presence of [3H]
glucosamine in serum-free medium were fractionated on the basis of their interaction with
WGA-Sepharose (2 mg/mL). The WGA-bound
glycoproteins from the two variants had molecular weights of 92,000, 56,000, and 42,000. None of these components was detected in the parent cell-line. A major WGA-binding
glycoprotein, which accounted for 37% of the total [3H]
glucosamine incorporated, was isolated from the spent medium of the parent mouse
melanoma cell-line. This
glycoprotein was apparently absent in the WGA-1-resistant variants.