TY - JOUR
T1 - Glycomics of a novel type-2 N-acetyllactosamine-specific lectin purified from the feather star, Oxycomanthus japonicus (Pelmatozoa
T2 - Crinoidea)
AU - Matsumoto, Ryo
AU - Shibata, Tomoko F.
AU - Kohtsuka, Hisanori
AU - Sekifuji, Mamoru
AU - Sugii, Natsuko
AU - Nakajima, Hiroaki
AU - Kojima, Noriaki
AU - Fujii, Yuki
AU - Kawsar, Sarkar M.A.
AU - Yasumitsu, Hidetaro
AU - Hamako, Jiharu
AU - Matsui, Taei
AU - Ozeki, Yasuhiro
N1 - Funding Information:
This work was supported in part by a Grant-In-Aid for scientific research (No. 1022580226 ) from the Japan Society for the Promotion of Science (JSPS) , Japanese Association for Marine Biology (JAMBIO, No. 22-02 ) from the Ministry of Education, Culture, Sports, Science and Technology (MEXT) Japan and Strategic Research Project (No. G2201 ) from Yokohama City University .
PY - 2011/4
Y1 - 2011/4
N2 - 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 kass and kdiss values of 1.4×10-6M-1s-1 and 3.1×10-3s-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.
AB - 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 kass and kdiss values of 1.4×10-6M-1s-1 and 3.1×10-3s-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.
KW - Crinoidea
KW - Echinodermata
KW - Feather star (Oxycomanthus japonicus)
KW - Frontal affinity chromatography
KW - Glycomics
KW - Lectin
KW - Pelmatozoa
KW - Type-2 N-acetyllactosamine
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U2 - 10.1016/j.cbpb.2010.12.004
DO - 10.1016/j.cbpb.2010.12.004
M3 - Article
C2 - 21176791
AN - SCOPUS:79951774682
VL - 158
SP - 266
EP - 273
JO - Comparative Biochemistry and Physiology - B Biochemistry and Molecular Biology
JF - Comparative Biochemistry and Physiology - B Biochemistry and Molecular Biology
SN - 1096-4959
IS - 4
ER -