Functional cooperation between the neural adhesion molecules L1 and N-CAM is carbohydrate dependent

The neural cell adhesion molecules L1 and N-CAM have been suggested to interact functionally by formation of a complex between the two molecules (Kadmon, G., A. Kowitz, P. Altevogt, and M. Schachner. 1990. J. Cell Biol. 110:193-208). To determine the molecular mechanisms underlying this functional cooperation, we have studied the contribution of carbohydrates to the association of the two molecules at the cell surface. Aggregation or adhesion between L1- and N-CAM-nositive neuroblastoma N2A cells was reduced when ne synthesis of complex and/or hybrid glycans was iodified by castanospermine. Fab fragments of polyional antibodies to L1 inhibited aggregation and adheion of castanospermine-treated cells almost completely, whereas untreated cells were inhibited by ∼50%. Fab fragments of polyclonal antibodies to N-CAM did not interfere with the interaction between astanospermine-treated cells, whereas they inhibited ggregation or adhesion of untreated cells by ∼50%. These findings indicate that cell interactions depending both on L1 and N-CAM ("assisted homophilic" binding) can be reduced to an L1-dominated interaction ("homophilic binding"). Treatment of cells with the carbohydrate synthesis inhibitor swainsonine did not modify cell aggregation in the absence or presence of antibodies compared with untreated cells, indicating that castanospermine-sensitive, but swainsonine-insensitive glycans are involved. To investigate whether the appropriate carbohydrate composition is required for an association of L1 and N-CAM in the surface membrane (cis-interaction) or between L1 on one side and L1 and N-CAM on the other side of interacting partner cells (trans-interaction), an L1-positive lymphoid tumor cell line was coaggregated with and adhered to neuroblastoma cells in the various combinations of castanospermine-treated and untreated cells. The results show that it is the m-interaction between L1 and N-CAM that depends on the appropriate carbohydrate structures.