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A highly sulfated 3-linked β-arabinan (Ab1) with arabinose in the pyranose form was obtained from green seaweed Codium vermilara (Bryopsidales). It comprised major amounts of units sulfated on C-2 and C-4 and constitutes the first polysaccharide of this type isolated in the pure form and fully characterized. Ab1 showed anticoagulant activity by global coagulation tests. Less sulfated arabinans obtained from the same seaweed have less or no activity. Ab1 exerts its activity through direct and indirect (antithrombin- and heparin cofactor II-mediated) inhibition of thrombin. Direct thrombin inhibition was studied in detail. By native PAGE, it was possible to detect formation of a complex between Ab1 and human thrombin (HT). Ab1 binding to HT was measured by fluorescence spectroscopy. CD spectra of the Ab1 complex suggested that ligand binding induced a small conformational change on HT. Ab1-thrombin interactions were studied by molecular dynamic simulations using the persulfated octasaccharide as model compound. Most carbohydrate-protein contacts would occur by interaction of sulfate groups with basic amino acid residues on the surface of the enzyme, more than 60% of them being performed by the exosite 2-composing residues. In these interactions, the sulfate groups on C-2 were shown to interact more intensely with the thrombin structure. In contrast, the disulfated oligosaccharide does not promote major conformational modifications at the catalytic site when complexed to exosite 1. These results show that this novel pyranosic sulfated arabinan Ab1 exerts its anticoagulant activity by a mechanism different from those found previously for other sulfated polysaccharides and glycosaminoglycans. © 2013 by The American Society for Biochemistry and Molecular Biology, Inc.


Documento: Artículo
Título:Anticoagulant activity of a unique sulfated pyranosic (1→3)-β-L- arabinan through direct interaction with thrombin
Autor:Fernández, P.V.; Quintana, I.; Cerezo, A.S.; Caramelo, J.J.; Pol-Fachin, L.; Verli, H.; Estevez, J.M.; Ciancia, M.
Filiación:Cátedra de Química de Biomoléculas, Departamento de Biología Aplicada y Alimentos, Universidad de Buenos Aires, Av. San Martín 4453, 1417 Buenos Aires, Argentina
Laboratorio de Hemostasia y Trombosis, Departamento de Química Biológica, Universidad de Buenos Aires, Pabellón 2, C1428EHA Buenos Aires, Argentina
Centro de Investigaciones en Hidratos de Carbono (CIHIDECAR), Departamento de Química Orgánica, Universidad de Buenos Aires, Pabellón 2, C1428EHA Buenos Aires, Argentina
Instituto de Investigaciones Bioquímicas de Buenos Aires (IIBBA), CONICET, Av. Patricias Argentinas 435, 1405 Buenos Aires, Argentina
Programa de Pos-Graduação em Biologia Celular e Molecular, Centro de Biotecnologia, Universidade Federal do Rio Grande do Sul, Porto Alegre, Rio Grande do Sul, Brazil
Faculdade de Farmácia, Universidade Federal do Rio Grande do Sul, 90610-000 Porto Alegre, Rio Grande do Sul, Brazil
Instituto de Fisiología, Biología Molecular y Neurociencias (IFIByNE-CONICET), Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Pabellón 2, C1428EHA Buenos Aires, Argentina
Palabras clave:Amino acid residues; Anticoagulant activities; Antithrombin; Arabinans; Catalytic sites; CD spectra; Conformational change; Direct interactions; Glycosaminoglycans; Green seaweed; Heparin cofactor; Ligand binding; Model compound; Octasaccharide; Pyranose; Sulfate groups; Sulfated polysaccharides; Thrombin inhibition; Amino acids; Coagulation; Computer simulation; Fluorescence spectroscopy; Oligosaccharides; Polysaccharides; Seaweed; Enzymes; algal extract; amino acid; arabinose; carbohydrate; heparin cofactor II; oligosaccharide; polysaccharide; protein; sulfated pyranosic beta arabinan; thrombin; unclassified drug; anticoagulation; article; blood clotting test; catalysis; circular dichroism; conformational transition; drug mechanism; drug structure; fluorescence spectroscopy; ligand binding; molecular dynamics; nonhuman; priority journal; protein modification; protein protein interaction; seaweed; simulation; Animals; Anticoagulants; Biophysics; Blood Coagulation; Carbohydrate Conformation; Cattle; Cell Wall; Circular Dichroism; Electrophoresis; Gas Chromatography-Mass Spectrometry; Humans; Kinetics; Magnetic Resonance Spectroscopy; Methylation; Models, Chemical; Molecular Conformation; Polysaccharides; Protein Binding; Pyrans; Seaweed; Spectroscopy, Fourier Transform Infrared; Thrombin; Caulerpales; Codium vermilara
Página de inicio:223
Página de fin:233
Título revista:Journal of Biological Chemistry
Título revista abreviado:J. Biol. Chem.
CAS:amino acid, 65072-01-7; arabinose, 147-81-9; heparin cofactor II, 81604-65-1; protein, 67254-75-5; thrombin, 869858-13-9, 9002-04-4; Anticoagulants; Polysaccharides; Pyrans; Thrombin,; araban, 11078-27-6; arabinan sulfate, 145378-97-8


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---------- APA ----------
Fernández, P.V., Quintana, I., Cerezo, A.S., Caramelo, J.J., Pol-Fachin, L., Verli, H., Estevez, J.M.,..., Ciancia, M. (2013) . Anticoagulant activity of a unique sulfated pyranosic (1→3)-β-L- arabinan through direct interaction with thrombin. Journal of Biological Chemistry, 288(1), 223-233.
---------- CHICAGO ----------
Fernández, P.V., Quintana, I., Cerezo, A.S., Caramelo, J.J., Pol-Fachin, L., Verli, H., et al. "Anticoagulant activity of a unique sulfated pyranosic (1→3)-β-L- arabinan through direct interaction with thrombin" . Journal of Biological Chemistry 288, no. 1 (2013) : 223-233.
---------- MLA ----------
Fernández, P.V., Quintana, I., Cerezo, A.S., Caramelo, J.J., Pol-Fachin, L., Verli, H., et al. "Anticoagulant activity of a unique sulfated pyranosic (1→3)-β-L- arabinan through direct interaction with thrombin" . Journal of Biological Chemistry, vol. 288, no. 1, 2013, pp. 223-233.
---------- VANCOUVER ----------
Fernández, P.V., Quintana, I., Cerezo, A.S., Caramelo, J.J., Pol-Fachin, L., Verli, H., et al. Anticoagulant activity of a unique sulfated pyranosic (1→3)-β-L- arabinan through direct interaction with thrombin. J. Biol. Chem. 2013;288(1):223-233.