RRC ID |
7199
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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.
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タイトル |
Glycomics of a novel type-2 N-acetyllactosamine-specific lectin purified from the feather star, Oxycomanthus japonicus (Pelmatozoa: Crinoidea).
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ジャーナル |
Comp Biochem Physiol B Biochem Mol Biol
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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.
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巻・号 |
158(4)
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ページ |
266-73
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公開日 |
2011-4-1
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DOI |
10.1016/j.cbpb.2010.12.004
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PII |
S1096-4959(10)00294-0
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PMID |
21176791
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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
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WOS 分野
|
ZOOLOGY
BIOCHEMISTRY & MOLECULAR BIOLOGY
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リソース情報 |
カタユウレイボヤ・(ニッポンウミシダ) |
O. japonicus |