Species-specific cell reaggregation in the marine sponge Microciona prolifera is mediated by an adhesion proteoglycan. Two interactions are involved in the process: a Ca2+-dependent homophilic binding between proteoglycan molecules and a Ca2+-independent binding between the molecule and cells. Both interactions are mediated by the glycan moieties of the proteoglycan. The interaction of the proteoglycan with itself has been characterized as a carbohydrate-carbohydrate interaction of multiple low affinity sites. The monoclonal antibodies Block 1 and Block 2 raised against the purified aggregation proteoglycan and selected for inhibition of aggregation bind to these glycans. In a previous report the structure, Pyr<(6)(4)>Gal beta 1-4GlcNAc beta 1-3Fuc STRUCTURE 1 was assigned to the oligosaccharide reacting with Block 1 antibody (Spillmann, D., Hard, K., Thomas-Gates, J., Vliegenthart, J. F. G., Misevic, G., Burger, M. M., and Finne, J. (1993) J. Biol, Chem. 268, 13378-13387), By the technique of attaching the water-soluble acid-degraded fragments to a lipid carrier for immunochemical detection and by chemical, enzymatic and spectroscopic methods the structure, GlcNAc beta 1-3Fuc\3SO(3) STRUCTURE 2 was assigned to the oligosaccharide reacting with the aggregation- blocking monoclonal antibody Block 2. The structure, Gal alpha 1-2Gal beta 1-4GlcNAc beta 1-3Fuc\3SO(3) was assigned to a major nonreactive oligosaccharide, which outlined the molecular requirements of antibody binding of the two aggregation-associated epitopes. These data demonstrate that two different functional sites with distinct structural characteristics and antibody reactivities are involved in the reaggregation of sponge cells, a model of carbohydrate carbohydrate-mediated cell interactions.
