Purification and characterization of an O-acetylsialic acid-specific lectin from a marine crab Cancer antennarius.
A sialic acid-binding lectin with high specificity for 9-O-acetyl- and 4-O-acetylsialic acids was purified from the hemolymph of the California coastal crab, Cancer antennarius, by affinity chromatography using bovine submaxillary mucin coupled to agarose. The binding specificity of the crab lectin distinguishes it from other known sialic acid-specific lectins from Limulus polyphemus and Limax flavus which show a broader range of specificity for sialic acids. The purified lectin is homogenous on sodium dodecyl sulfate-polyacrylamide electropherograms with a subunit molecular weight of about 36 kDa. The specificity of the lectin for O-acetylsialic acids appears to account for the fact that it agglutinates mouse, rat, rabbit, and horse erythrocytes, which contain O-acetylsialic acids on cell surface glycoconjugates, but not human monkey, sheep, goat, and chicken erythrocytes which contain only NeuAc or N-glycolylneuraminic acid (NeuGc). This conclusion was supported by the potent inhibition of hemagglutination by bovine and equine submaxillary mucins which contain 9(7,8)-O-acetyl- and 4-O-acetylsialic acids, respectively, and also by free 9-O-acetyl-N-acetylneuraminic acid (9-O-Ac-NeuAc) and 4-O-Ac-NeuAc relative to NeuAc and NeuGc. Further support for the role of O-Ac-sialic acids in hemagglutination of erythrocytes was obtained by enzymatic modification of human erythrocytes. Sialidase-treated erythrocytes were resialylated with purified sialyltransferases and various CMP-sialic acid donor substrates to contain NeuAc or NeuGc or 9-O-Ac-NeuAc in the Sia alpha 2,3Gal or Sia alpha 2,6Gal linkages. Cells resialylated to contain NeuAc or NeuGc were not agglutinated, but cells resialylated to contain 9-O-Ac-NeuAc were agglutinated with high titer, comparable to that of mice or horse erythrocytes.
D000818:Animals; D003386:Brachyura / Q000032:analysis*; D002846:Chromatography, Affinity; D004912:Erythrocytes / Q000378:metabolism; D006384:Hemagglutination; D006801:Humans; D037102:Lectins / Q000302:isolation & purification* / Q000502:physiology; D051379:Mice; D009077:Mucins / Q000494:pharmacology; D011817:Rabbits; D051381:Rats; D012794:Sialic Acids / Q000032:analysis / Q000378:metabolism; D013045:Species Specificity
D037102:Lectins; D009077:Mucins; D012794:Sialic Acids; C041901:4-O-acetyl-N-acetylneuraminic acid; C046053:8-O-acetyl-N-acetylneuraminic acid; C025676:9-O-acetyl-N-acetylneuraminic acid