Purification and characterization of a new lectin from the hard roe of skipjack tuna, Katsuwonus pelamis.
Fish eggs are known as a rich source of lectins. In this study we purified and characterized a lectin from unfertilized Katsuwonus pelamis hard roe. K. pelamis lectin (KPL) was purified by separation into two fractions above and below the molecular weight of 10kDa using ultramembrane, gel filtration on a Sephadex G-100, and affinity chromatography on an asialofetuin-Sepharose 4B. KPL is a glycoprotein of 140kDa, composed mainly of aspartic acid, glycine, phenylalanine, glutamic acid, threonine and serine residues. Analysis of the carbohydrate composition by gas-liquid chromatography indicated that carbohydrates constituted 14% of the total weight and this 14% is comprised of mannose, galactose, N-acetyl-D-glucosamine, N-acetyl-D-galactosamine, fucose, arabinose and sialic acid. The lectin is comprised of four subunits. These subunits have a molecular mass corresponding to 35kDa. KPL specifically agglutinated human blood type A erythrocytes and, in a hemagglutination inhibitory test, the potent inhibitors were D-galactose, lactose, lactosamine, asialofetuin, N-acetyl-D-galactosamine, O-serinyl-2-acetamido-2-deoxy-alpha-D-galactopyranoside and O-serinyl-2-acetamido-2-deoxy-beta-D-galactopyranoside (O-serinyl-beta-D-GalNAc). The first 10 residues of the N-terminal region were determined as PVELCDAKCT. Furthermore it was determined that the hemagglutinating activity of KPL was dependent on divalent metal cations and that the optimum activity of KPL was exhibited at 40 degrees C and pH 6.0-8.5 in the presence of Ca2+.
Jung
Won-Kyo
WK
Department of Chemistry, Pukyong National University, 608-737, Busan, South Korea.
Park
Pyo-Jam
PJ
Kim
Se-Kwon
SK
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002241:Carbohydrates / Q000032:analysis / Q000494:pharmacology; D002412:Cations / Q000378:metabolism / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D006023:Glycoproteins / Q000494:pharmacology; D006384:Hemagglutination / Q000187:drug effects; D006385:Hemagglutination Inhibition Tests; D006801:Humans; D006863:Hydrogen-Ion Concentration; D037102:Lectins / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000302:isolation & purification* / Q000494:pharmacology; D008970:Molecular Weight; D010063:Ovum / Q000737:chemistry; D013696:Temperature; D014413:Tuna
D000596:Amino Acids; D002241:Carbohydrates; D002412:Cations; D006023:Glycoproteins; D037102:Lectins