Isolation, purification, characterization and glycan-binding profile of a d-galactoside specific lectin from the marine sponge, Halichondria okadai.
A lectin recognizing both Galbeta1-3GlcNAc and Galbeta1-4GlcNAc was purified from the demosponge Halichondria okadai by lactosyl-agarose affinity chromatography. The molecular mass of the lectin was determined to be 30 kDa by SDS-PAGE under reducing and non-reducing conditions and 60 kDa by gel permeation chromatography. The pI value of the lectin was 6.7. It was found to agglutinate trypsinized and glutaraldehyde-fixed rabbit and human erythrocytes in the presence and absence of divalent cations. The hemagglutinating activity by the lectin was inhibited by d-galactose, methyl-d-galactopyranoside, N-acetyl-d-galactosamine, methyl-N-acetyl-d-galactosaminide, lactose, melibiose, and asialofetuin. The K(d) of the lectin against p-nitrophenyl-beta-lactoside was determined to be 2.76x10(-5) M and its glycan-binding profile given by frontal affinity chromatography was shown to be similar to many other known galectins. Partial primary structure analysis of 7 peptides by cleavage with lysyl endopeptidase indicated that one of the peptides showed significant similarity with galectin purified from the sponge Geodia cydonium.
Laboratory of Marine Biochemistry, Department of Environmental Biosciences, International Graduate School of Arts and Sciences, Yokohama City University, 22-2 Seto, Kanazawa-ku, Yokohama 236-0027, Japan.
Sarkar M A
SM
Kawsar
Yuki
Y
Fujii
Ryo
R
Matsumoto
Takayuki
T
Ichikawa
Hiroaki
H
Tateno
Jun
J
Hirabayashi
Hidetaro
H
Yasumitsu
Chikaku
C
Dogasaki
Masahiro
M
Hosono
Kazuo
K
Nitta
Jiharu
J
Hamako
Taei
T
Matsui
Yasuhiro
Y
Ozeki
D000116:Acetylgalactosamine / Q000494:pharmacology; D000818:Animals; D001212:Asialoglycoproteins / Q000494:pharmacology; D002240:Carbohydrate Sequence; D002413:Cations, Divalent / Q000494:pharmacology; D002846:Chromatography, Affinity; D003619:Dansyl Compounds / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D060748:Fetuins; D005688:Galactosamine / Q000031:analogs & derivatives / Q000494:pharmacology; D005690:Galactose / Q000494:pharmacology; D037161:Galectins / Q000302:isolation & purification* / Q000378:metabolism* / Q000494:pharmacology; D006027:Glycosides / Q000378:metabolism; D006384:Hemagglutination / Q000187:drug effects; D006801:Humans; D007525:Isoelectric Focusing; D007785:Lactose / Q000494:pharmacology; D008553:Melibiose / Q000494:pharmacology; D008756:Methylgalactosides / Q000494:pharmacology; D008969:Molecular Sequence Data; D011161:Porifera / Q000378:metabolism*; D011817:Rabbits; D000509:alpha-Fetoproteins / Q000494:pharmacology
D001212:Asialoglycoproteins; D002413:Cations, Divalent; D003619:Dansyl Compounds; D060748:Fetuins; D037161:Galectins; D006027:Glycosides; D008756:Methylgalactosides; D000509:alpha-Fetoproteins; C017029:asialofetuin; C008245:lactosides; C079425:methyl-N-dansylgalactosaminide; D005688:Galactosamine; C057031:methyl 4-O-galactopyranosylgalactopyranoside; D008553:Melibiose; D007785:Lactose; D000116:Acetylgalactosamine; D005690:Galactose