A single step purification of a sialic acid binding lectin (AchatininH) from Achatina fulica snail.
A sialic acid binding lectin, AchatininH, was purified in single step from the hemolymph of the land snail, Achatina fulica, by the affinity chromatography on sheep submaxillary mucin coupled to Sepharose 4B. The yield of the lectin was found to be 3 mg from 100 ml of hemolymph. The homogeneity of the lectin was established by alkaline gel electrophoresis, immunodiffusion, immunoelectrophoresis and analytical isoelectrophoresis. The molecular weight of the native protein was 242,000, having identical subunits of Mr 15,000. The lectin agglutinated rabbit erythrocytes in the presence of Ca2+. The inhibition study clearly suggests that the binding site of the lectin recognizes sialic acid as the immunodominant sugar. This was further confirmed by the observation that there was a marked decrease of agglutinating activity of the lectin with neuraminidase treated rabbit erythrocytes and asialofetuin was unable to inhibit the activity of AchatininH. Among the inhibitors used the glycoconjugate containing alpha 2----6 linkages of N-acetylneuraminic acid with subterminal galactopyranose or 2-acetamido-2-deoxy-galactopyranose residue was found to be better inhibitor than that containing alpha 2----3 linkages of N-acetyl neuraminic acid. Besides that sialoglycoprotein containing both N and O type of glycosidic linkages plays an important role in binding with the lectin. Fetuin was found to be the best inhibitor.
D000818:Animals; D002118:Calcium / Q000494:pharmacology; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D002846:Chromatography, Affinity; D006384:Hemagglutination / Q000187:drug effects; D006458:Hemolymph / Q000032:analysis; D037102:Lectins / Q000037:antagonists & inhibitors / Q000302:isolation & purification*; D008970:Molecular Weight; D009439:Neuraminidase / Q000494:pharmacology; D011817:Rabbits; D012908:Snails / Q000032:analysis*; D000509:alpha-Fetoproteins / Q000494:pharmacology
D037102:Lectins; C050704:achatinin(H); D000509:alpha-Fetoproteins; D009439:Neuraminidase; D002118:Calcium