1. Mannan-binding lectin of the sea urchin Strongylocentrotus nudus.
- Author
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Bulgakov AA, Eliseikina MG, Kovalchuk SN, Petrova IY, Likhatskaya GN, Shamshurina EV, and Rasskazov VA
- Subjects
- Amino Acid Sequence, Animals, Base Sequence, Calcium metabolism, Chromatography, Affinity, Chromatography, Agarose, Chromatography, Gel, Chromatography, Ion Exchange, Cross Reactions, Dimerization, Electrophoresis, Polyacrylamide Gel, Hemagglutination Inhibition Tests, Humans, Hydrogen-Ion Concentration, Immunohistochemistry, Mannose-Binding Lectin isolation & purification, Molecular Sequence Data, Sequence Analysis, DNA, Species Specificity, Strongylocentrotus immunology, Temperature, Mannose-Binding Lectin chemistry, Mannose-Binding Lectin genetics, Models, Molecular, Protein Conformation, Strongylocentrotus genetics
- Abstract
A novel lectin specific to low-branched mannans (MBL-SN) was isolated from coelomic plasma of the sea urchin Strongylocentrotus nudus by combining anion-exchange liquid chromatography on DEAE Toyopearl 650 M, affinity chromatography on mannan-Sepharose and gel filtration on the Sephacryl S-200. The molecular mass of MBL-SN was estimated by sodium dodecyl sulphate polyacrylamide gel electrophoresis under non-reducing conditions to be about 34 kDa. MBL-SN was shown to be a dimer with two identical subunits of about 17 kDa. The native MBL-SN exists as a tetramer. The physico-chemical properties of MBL-SN indicate that it belongs to C-type mannan-binding lectins. The cDNA encoding MBL-SN was cloned from the total cDNA of S. nudus coelomocytes and encodes a 17-kDa protein of 144 amino acid residues that contains a single carbohydrate-recognition domain of C-type lectins. Prediction of the MBL-SN tertiary structure using comparative modelling revealed that MBL-SN is an α/β-protein with eight β-strands and two α-helices. Comparison of the MBL-SN model with available three-dimensional structures of C-type lectins revealed that they share a common fold pattern.
- Published
- 2013
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