title: Glycomics of a novel type-2 N-acetyllactosamine-specific lectin purified from the feather star, Oxycomanthus japonicus (Pelmatozoa: Crinoidea).
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.
authors.affiliation: Laboratory of Glycobiology and Marine Biochemistry, Department of Genome System Sciences, Graduate School of NanoBiosciences, Yokohama City University, 22-2 Seto, Kanazawa-ku, Yokohama 236-0027, Japan.
authors.forename: Ryo
authors.initials: R
authors.lastname: Matsumoto
authors.forename: Tomoko F
authors.initials: TF
authors.lastname: Shibata
authors.forename: Hisanori
authors.initials: H
authors.lastname: Kohtsuka
authors.forename: Mamoru
authors.initials: M
authors.lastname: Sekifuji
authors.forename: Natsuko
authors.initials: N
authors.lastname: Sugii
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Nakajima
authors.forename: Noriaki
authors.initials: N
authors.lastname: Kojima
authors.forename: Yuki
authors.initials: Y
authors.lastname: Fujii
authors.forename: Sarkar M A
authors.initials: SM
authors.lastname: Kawsar
authors.forename: Hidetaro
authors.initials: H
authors.lastname: Yasumitsu
authors.forename: Jiharu
authors.initials: J
authors.lastname: Hamako
authors.forename: Taei
authors.initials: T
authors.lastname: Matsui
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Ozeki
mesh_terms: D000606:Amino Sugars / Q000378:metabolism*; D000818:Animals; D002413:Cations, Divalent / Q000737:chemistry; D004187:Disaccharides / Q000378:metabolism; D004447:Echinodermata / Q000737:chemistry*; D054794:Glycomics; D006863:Hydrogen-Ion Concentration; D037102:Lectins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D008970:Molecular Weight; D009005:Monosaccharides / Q000378:metabolism; D011134:Polysaccharides / Q000378:metabolism; D020349:Surface Plasmon Resonance; D013696:Temperature
chemical_list: D000606:Amino Sugars; D002413:Cations, Divalent; D004187:Disaccharides; D037102:Lectins; D009005:Monosaccharides; D011134:Polysaccharides; C000458:N-acetyllactosamine