Purification and characterization of two lectins from the sea cucumber Stichopus japonicus.
Two Ca(2+)-dependent lectins were purified from the sea cucumber Stichopus japonicus by affinity chromatography on lactosyl-Sepharose 4B and ion-exchange chromatography on Q-Sepharose. Their molecular masses were estimated to be 13kDa (SJL-I) and 15 kDa (SJL-II) on SDS-PAGE. SJL-I agglutinated rabbit erythrocytes as well as human A, B, and O-type erythrocytes, but SJL-II agglutinated only rabbit erythrocytes. Hemagglutination by SJL-I was competitively inhibited by N-acetyl-D-galactosamine and galactose-containing carbohydrates. On the other hand, only lactose, melibiose, and raffinose gave weak inhibition of hemagglutination by SJL-II, suggesting that SJL-II may have high specificity for particular complex carbohydrate(s) on the surface of rabbit erythrocytes. SJL-II was activated at ten times lower Ca2+ concentration than SJL-I. Both lectins lost activity in acidic pH, while SJL-I appeared more stable down to pH 4.5.
Hatakeyama
T
T
Laboratory of Biochemistry, Faculty of Agriculture, Kyushu University, Fukuoka, Japan.
Himeshima
T
T
Komatsu
A
A
Yamasaki
N
N
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002118:Calcium / Q000494:pharmacology; D002240:Carbohydrate Sequence; D002846:Chromatography, Affinity; D002852:Chromatography, Ion Exchange; D006384:Hemagglutination; D006801:Humans; D006863:Hydrogen-Ion Concentration; D037102:Lectins / Q000737:chemistry / Q000302:isolation & purification* / Q000494:pharmacology; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000494:pharmacology; D011817:Rabbits; D012616:Sea Cucumbers / Q000737:chemistry*
D000596:Amino Acids; D037102:Lectins; D009844:Oligosaccharides; D002118:Calcium