期刊
COMPARATIVE BIOCHEMISTRY AND PHYSIOLOGY B-BIOCHEMISTRY & MOLECULAR BIOLOGY
卷 152, 期 4, 页码 382-389出版社
ELSEVIER SCIENCE INC
DOI: 10.1016/j.cbpb.2009.01.009
关键词
Frontal affinity chromatography; Galactose-binding lectin; Lugworm (Perinereis nuntia var. vallata); Primary structure; Surface plasmon resonance
资金
- JSPS (Japan Society for the Promotion of Science) [0760110001]
- Yokohama City University [K20002]
- Ministry of Education, Culture, Science, Sports, and Technology (MEXT), Japan.
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 lactosylagarose 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 (Gal beta 1-3GlcNAc) and type 2 (Gal beta 1-4GlcNAc) lactosamine. The surface plasmon resonance study of the lectin against asialofetuin showed the k(ass) and k(diss) values are 5.14 x 10(4) M-1 s(-1) and 2.9 x 10(-3) s(-1), respectively. The partial primary structure of the lectin reveals 182 amino acids with novel sequence. (C) 2009 Elsevier Inc. All rights reserved.
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