RRC ID 7199
著者 Matsumoto R, Shibata TF, Kohtsuka H, Sekifuji M, Sugii N, Nakajima H, Kojima N, Fujii Y, Kawsar SM, Yasumitsu H, Hamako J, Matsui T, Ozeki Y.
タイトル Glycomics of a novel type-2 N-acetyllactosamine-specific lectin purified from the feather star, Oxycomanthus japonicus (Pelmatozoa: Crinoidea).
ジャーナル Comp Biochem Physiol B Biochem Mol Biol
Abstract A lectin - designated OXYL for the purposes of this study that strongly recognizes complex-type oligosaccharides of serum glycoproteins - was purified from a crinoid, the feather star Oxycomanthus japonicus, the most basal group among extant echinoderms. OXYL was purified through a combination of anion-exchange and affinity chromatography using Q-sepharose and fetuin-sepharose gel, respectively. Lectin was determined to be a 14-kDa polypeptide by sodium dodecyl sulphate-polyacrylamide gel electrophoresis under reducing conditions. However, 14-kDa and 28-kDa bands appeared in the same proportion under non-reducing conditions. Gel permeation chromatography showed a 54-kDa peak, suggesting that lectin consists of four 14-kDa subunits. Divalent cations were not indicated, and stable haemagglutination activity was demonstrated at pH 4-12 and temperatures below 60°C. Surface plasmon resonance analysis of OXYL against fetuin showed k(ass) and k(diss) values of 1.4×10(-6)M(-1)s(-1) and 3.1×10(-3)s(-1), respectively, indicating that it has a strong binding affinity to the glycoprotein as lectin. Frontal affinity chromatography using 25 types of prydylamine-conjugated glycans indicated that OXYL specifically recognizes multi-antennary complex-type oligosaccharides containing type-2 N-acetyllactosamines (Galβ1-4GlcNAc) if α2-3-linked sialic acid is linked at the non-reducing terminal. However, type-1 N-acetyllactosamine (Galβ1-3GlcNAc) chains and α2-6-linked sialic acids were never recognized by OXYL. This profiling study showed that OXYL essentially recognizes β1-4-linkage at C-1 position and free OH group at C-6 position of Gal in addition to the conservation of N-acetyl groups at C-2 position and free OH groups at C-3 position of GlcNAc in N-acetyllactosamine. This is the first report on glycomics on a lectin purified from an echinoderm belonging to the subphylum Pelmatozoa.
巻・号 158(4)
ページ 266-73
公開日 2011-4-1
DOI 10.1016/j.cbpb.2010.12.004
PII S1096-4959(10)00294-0
PMID 21176791
MeSH Amino Sugars / metabolism* Animals Cations, Divalent / chemistry Disaccharides / metabolism Echinodermata / chemistry* Glycomics Hydrogen-Ion Concentration Lectins / chemistry Lectins / isolation & purification Lectins / metabolism* Molecular Weight Monosaccharides / metabolism Polysaccharides / metabolism Surface Plasmon Resonance Temperature
IF 2.219
引用数 9
WOS 分野 ZOOLOGY BIOCHEMISTRY & MOLECULAR BIOLOGY
リソース情報
カタユウレイボヤ・(ニッポンウミシダ) O. japonicus