Structural characterization of a rhamnose-binding glycoprotein (lectin) from Spanish mackerel (Scomberomorous niphonius) eggs |
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Authors: | Takatomo Terada Yasuharu Watanabe Hiroaki Tateno Takako Naganuma Tomohisa Ogawa Koji Muramoto Hisao Kamiya |
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Affiliation: | 1. Department of Biomolecular Science, Graduate School of Life Sciences, Tohoku University, Sendai 981-8555, Japan;2. School of Fisheries Sciences, Kitasato University, Ohfunato, Iwate 022-0101, Japan |
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Abstract: | A rhamnose-binding glycoprotein (lectin), named SML, was isolated from the eggs of Spanish mackerel (Scomberomorous niphonius) by affinity and ion-exchange chromatographies. SML was composed of a non-covalently linked homodimer. The SML subunit was composed of 201 amino acid residues with two tandemly repeated domains, and contained 8 half-Cys residues in each domain, which is highly homologous to the N-terminal lectin domain of calcium-independent α-latrotoxin receptor in mammalian brains. Each domain has the same disulfide bonding pattern; Cys10–Cys40, Cys20–Cys99, Cys54–Cys86 and Cys67–Cys73 were located in the N-terminal domain, and Cys108–Cys138, Cys117–Cys195, Cys152–Cys182 and Cys163–Cys169 were in the C-terminal domain. SML was N-glycosylated at Asn168 in the C-terminal domain. The structure of the sugar chain was determined to be NeuAc-Galβ1-4GlcNAcβ1-2Manα1-6-(NeuAc-Galβ1-4GlcNAcβ1-2Manα1-3)Manβ1-4GlcNAcβ1-4GlcNAc-Asn. |
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Keywords: | BCA, bincinchonic acid BSA, bovine serum albumin CAM, S-carboxamidomethylated CRD, carbohydrate-recognition domain CSL, chum salmon egg lectin DHBA, 2,5-dihydroxybenzoic acid DMB, 1,2-diamino-4,5-methylene dioxybenzene 2D, two dimensional kDa, kilodalton MALDI-TOF, matrix-assisted laser desorption ionization time of flight 2-ME, 2-mercaptoethanol NeuAc, N-acetylneuraminic acid NeuGc, N-glycolylneuraminic acid PA, pyridyl amine PE, pyridyl ethyl RBL, rhamnose-binding lectin RCLD, RBL CRD-like domain RP-HPLC, reversed-phase high-performance liquid chromatography SDS-PAGE, sodium dodecyl sulfate polyacrylamide gel electrophoresis SML, Spanish mackerel egg lectin STL, steelhead tout egg lectin SUEL, sea urchin egg lectin TBS, 20  mM Tris&ndash HCl buffer (pH 8.0) containing 0.5 M NaCl TFA, trifluoroacetic acid WCL, white-spotted charr egg lectin |
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