Glycomics of a novel type-2 N-acetyllactosamine-specific lectin purified from the feather star, Oxycomanthus japonicus (Pelmatozoa: Crinoidea)

Ryo Matsumoto, Tomoko F. Shibata, Hisanori Kohtsuka, Mamoru Sekifuji, Natsuko Sugii, Hiroaki Nakajima, Noriaki Kojima, Yuki Fujii, Sarkar M.A. Kawsar, Hidetaro Yasumitsu, Jiharu Hamako, Taei Matsui, Yasuhiro Ozeki

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13 Citations (Scopus)

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 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.

Original languageEnglish
Pages (from-to)266-273
Number of pages8
JournalComparative Biochemistry and Physiology - B Biochemistry and Molecular Biology
Volume158
Issue number4
DOIs
Publication statusPublished - 2011 Apr

Keywords

  • Crinoidea
  • Echinodermata
  • Feather star (Oxycomanthus japonicus)
  • Frontal affinity chromatography
  • Glycomics
  • Lectin
  • Pelmatozoa
  • Type-2 N-acetyllactosamine

ASJC Scopus subject areas

  • Biochemistry
  • Physiology
  • Molecular Biology

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    Matsumoto, R., Shibata, T. F., Kohtsuka, H., Sekifuji, M., Sugii, N., Nakajima, H., Kojima, N., Fujii, Y., Kawsar, S. M. A., Yasumitsu, H., Hamako, J., Matsui, T., & Ozeki, Y. (2011). Glycomics of a novel type-2 N-acetyllactosamine-specific lectin purified from the feather star, Oxycomanthus japonicus (Pelmatozoa: Crinoidea). Comparative Biochemistry and Physiology - B Biochemistry and Molecular Biology, 158(4), 266-273. https://doi.org/10.1016/j.cbpb.2010.12.004