Physicochemical and functional characterization of the polymerization process of the Geodia cydonium lectin.
The extracellularly localized, galactose-specific lectin from the sponge Geodia cydonium binds at one class of sites, 40 mol Ca2+/mol lectin with an association constant (Ka) of 0.3 X 10(6)M-1. Stoichiometric calculations reveal that in the extracellular milieu 22 mol Ca2+ (maximum) are complexed per mol lectin. Binding of Ca2+ to the lectin increases its apparent Mr from 44000 to 56000 (electrophoretic determination) or from 36500 to 53500 (high-pressure liquid gel chromatographical determination); the s20, w increases from 4.3 S to 4.5 S if Ca2+ is added to the lectin. In the presence of Ca2+ the lectin undergoes a conformational change perhaps by expanding the carbohydrate side chains which are terminated by galactose. Subsequently the lectin molecules polymerize to large three-dimensional clumps (diameter up to 8 micron). Turbidimetric studies reveal an inhibition of the lectin polymerization by lactose. The Ka of the lectin-lectin polymerization rises from 0.9 X 10(6)M-1 to 14.0 X 10(6)M-1 after increasing the Ca2+ concentration (from 1 microM to 100 microM). Parallel with this increase in affinity, the Ka value of the lectin-aggregation factor binding drops from 41.2 X 10(6)M-1 (1 microM Ca2+) to 1.3 X 10(6)M-1 (100 microM Ca2+). In the absence of Ca2+, the Geodia lectin forms 1-10-micron two-dimensional sheets in the presence of homologous glycoconjugates. Cell binding experiments with polyacrylamide gels, containing covalently bound galactose, show that both homologous (Geodia cydonium) and heterologous cells (L5178y) bind with a higher affinity to the lectin-polymer matrix than to the lectin-monomer one. These data suggest that lectin-polymer structures, together with lectin-glycoconjugate associates, are components of the cell-substrate adhesion system(s) of sponges in vivo.
D000818:Animals; D001704:Biopolymers / Q000378:metabolism; D002118:Calcium / Q000378:metabolism; D002448:Cell Adhesion; D055598:Chemical Phenomena; D002621:Chemistry; D037102:Lectins / Q000032:analysis*; D007940:Leukemia L5178; D051379:Mice; D008854:Microscopy, Electron; D008970:Molecular Weight; D009391:Nephelometry and Turbidimetry; D011161:Porifera / Q000378:metabolism*; D011485:Protein Binding
D001704:Biopolymers; D037102:Lectins; D002118:Calcium