Polysaccharide structural features that are critical for the binding of sulfated fucans to bindin, the adhesive protein from sea urchin sperm.
We have investigated the structural features of sulfated fucose-containing polysaccharides which are responsible for their selective binding to Strongylocentrotus purpuratus bindin. The data presented demonstrate that the sulfate esters and a molecular weight in excess of approximately 15,000 are required for high affinity binding of the fucans to bindin. Desulfation destroys the binding activity of the fucans, which can be fully restored by chemical resulfation. Fucan fragments of an average molecular weight of 15,000 were nearly as active as the starting material (Mr 10(6)). The observed IC50 value for fragments of Mr congruent to 10,000 and Mr congruent to 5,000 were 1 and 2 orders of magnitude higher, respectively. The binding of fucoidan to bindin is stable in high salt (50% at 1.2 M NaCl) whereas the binding of fucoidan to DEAE-cellulose or polylysine is inhibited by the concentrations of salt normally found in sea water (50% at 0.2 and 0.5 M NaCl, respectively). This result suggests that the binding mechanism is not a simple ionic interaction and that hydrogen bonding and cooperativity may also be important determinants of the binding mechanism. We also found that polyvinyl sulfate binds to bindin with high affinity and inhibits the bindin-mediated agglutination of sea urchin eggs. The results of these investigations suggest that the spatial orientation of the sulfate esters plays a critical role in determining the selectivity of sulfated polysaccharide binding and that the polysaccharide backbone does not play a direct role in the binding mechanism.
DeAngelis
P L
PL
Department of Molecular Biology and Biochemistry, University of California, Irvine 92717.
Glabe
C G
CG
D000818:Animals; D005260:Female; D006023:Glycoproteins / Q000378:metabolism*; D008297:Male; D008970:Molecular Weight; D011134:Polysaccharides / Q000302:isolation & purification / Q000378:metabolism*; D011485:Protein Binding; D011956:Receptors, Cell Surface; D012617:Sea Urchins; D013084:Sperm-Ovum Interactions; D013094:Spermatozoa / Q000502:physiology; D013329:Structure-Activity Relationship
D006023:Glycoproteins; D011134:Polysaccharides; D011956:Receptors, Cell Surface; C035027:egg surface sperm receptor; C007789:fucoidan