Glycan-binding profile of a D-galactose binding lectin purified from the annelid, Perinereis nuntia ver. vallata.
A lectin recognizing D-galactose was purified from the pacific annelid Perinereis nuntia ver. vallata (Polychaeta) by affinity chromatography. Hemagglutinating activity, with a very low titer suggesting the presence of lectin appeared in the supernatant from the homogenization of body with Tris-buffered saline. However, dialyzed supernatant from the precipitate homogenized by galactose in the buffer revealed strong hemagglutinating activity against human erythrocytes. The crude supernatant was applied onto lactosyl-agarose column, and only the supernatant eluted from precipitate with galactose was obtained a galactose-binding lectin with 32 kDa polypeptide was obtained from the supernatant of the precipitate, extracted in presence of galactose. It suggests that the lectin tightly binds with glycoconjugate as endogenous ligand(s) in the tissue. Hemagglutinating activity against trypsinized and glutaraldehyde-fixed human erythrocytes was specifically inhibited by D-galactose, N-acetyl-D-galactosamine, lactose, melibiose, and asialofetuin. Glycan-binding profile of the lectin analyzed by frontal affinity chromatography shows that the lectin recognizes branched complex type N-linked oligosaccharides and both type 1 (Galbeta1-3GlcNAc) and type 2 (Galbeta1-4GlcNAc) lactosamine. The surface plasmon resonance study of the lectin against asialofetuin showed the k(ass) and k(diss) values are 5.14x10(4) M(-1) s(-1) and 2.9x10(-3) s(-1), respectively. The partial primary structure of the lectin reveals 182 amino acids with novel sequence.
Kawsar
Sarkar M A
SM
Department of Environmental Biosciences, Yokohama City University, Seto, Kanazawa-ku, Japan.
Takeuchi
Tomoharu
T
Kasai
Ken-ichi
K
Fujii
Yuki
Y
Matsumoto
Ryo
R
Yasumitsu
Hidetaro
H
Ozeki
Yasuhiro
Y
D000818:Animals; D001665:Binding Sites; D004912:Erythrocytes / Q000276:immunology; D005690:Galactose / Q000737:chemistry / Q000378:metabolism*; D006385:Hemagglutination Inhibition Tests; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D037102:Lectins / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification* / Q000378:metabolism*; D011077:Polychaeta / Q000737:chemistry*; D011134:Polysaccharides / Q000737:chemistry* / Q000378:metabolism*; D013696:Temperature
D037102:Lectins; D011134:Polysaccharides; D005690:Galactose