Journal of Cell Science, Vol 38, Issue 1 315-329, Copyright © 1979 by Company of Biologists

JOURNAL ARTICLES

The interaction of lectins with the surface of differentiating erythroleukaemic cells

RC Hunt and LM Marshall

Friend erythroleukaemic cells can be induced to mature along the erythroid differentiation pathway when an inducing agent such as dimethyl sulphoxide is included in the medium. In the absence of the inducing agent, the 707B line of Friend erythroleukaemic cells is highly agglutinable by the lectins concanavalin A or wheat germ agglutinin. However, 48 h after the induction of differentiation, there is a marked decrease in the agglutination of the cells in the presence of either lectin. This suggests that early in differentiation a change occurs in the cell membrane preceding the onset of globin synthesis which starts approximately 72 h after induction. The change in agglutination by concanavalin A also occurs in the presence of reagents which do not induce haemoglobin synthesis in the 707B line of Friend erythroleukaemic cells but which are able to stimulate the synthesis of this protein in other erythroleukaemic cell lines. The reduction in the agglutinability of the differentiating cells does not seem to result from a reduction in the number of concanavalin A receptors on the cells, nor does it reflect a change in the clustered distribution of concanavalin A receptors in the differentiating cells. Both the control and dimethyl sulphoxide-induced cells show a similar patchy distribution of ferritin-labelled concanavalin A when examined by electron microscopy. Polyacrylamide gel electrophoresis shows little change in the total pattern of protein synthesis by control and differentiating cells when pulse-labelled with [35S] methionine. However, use of 125I-labelled concanavalin A to stain polyacrylamide gels, on which the total proteins of control and differentiating cells had been separated, revealed a profound change in the composition of the concanavalin A-binding proteins. The control, undifferentiated cells contained eleven or more classes of concanavalin A-binding glycoproteins, many of which stained to a lesser degree as the cell density increased. After the onset of differentiation, 2 new concanavalin A-binding glycoproteins appeared within 48 h. One of these proteins has a molecular weight in excess of 180 000 while the other migrated with an apparent molecular weight of approximately 100 000. After erythroid differentiation had progressed for 120 h, these newly synthesized glycoproteins became the major concanavalin A-binding proteins of the erythroleukaemic cells.