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.
Kawsar
Sarkar M A
SM
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.
Fujii
Yuki
Y
Matsumoto
Ryo
R
Ichikawa
Takayuki
T
Tateno
Hiroaki
H
Hirabayashi
Jun
J
Yasumitsu
Hidetaro
H
Dogasaki
Chikaku
C
Hosono
Masahiro
M
Nitta
Kazuo
K
Hamako
Jiharu
J
Matsui
Taei
T
Ozeki
Yasuhiro
Y
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