Anticoagulant activity of a unique sulfated pyranosic (1→3)-β-L- arabinan through direct interaction with thrombin

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 chara...

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Autores principales:	Fernández, Paula Virginia, Quintana, Irene Luisa, Cerezo, Alberto Saúl, Estevez, Jose Manuel, Ciancia, Marina
Publicado:	2013
Materias:	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 Protein Binding Pyrans Spectroscopy, Fourier Transform Infrared Thrombin Caulerpales Codium vermilara
Acceso en línea:	https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00219258_v288_n1_p223_Fernandez http://hdl.handle.net/20.500.12110/paper_00219258_v288_n1_p223_Fernandez
Aporte de:	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) de Universidad de Buenos Aires

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spelling	paper:paper_00219258_v288_n1_p223_Fernandez2023-06-08T14:43:34Z Anticoagulant activity of a unique sulfated pyranosic (1→3)-β-L- arabinan through direct interaction with thrombin Fernández, Paula Virginia Quintana, Irene Luisa Cerezo, Alberto Saúl Estevez, Jose Manuel Ciancia, Marina 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 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. Fil:Fernández, P.V. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Quintana, I. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Cerezo, A.S. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Estevez, J.M. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Ciancia, M. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. 2013 https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00219258_v288_n1_p223_Fernandez http://hdl.handle.net/20.500.12110/paper_00219258_v288_n1_p223_Fernandez
institution	Universidad de Buenos Aires
institution_str	I-28
repository_str	R-134
collection	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA)
topic	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
spellingShingle	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 Fernández, Paula Virginia Quintana, Irene Luisa Cerezo, Alberto Saúl Estevez, Jose Manuel Ciancia, Marina Anticoagulant activity of a unique sulfated pyranosic (1→3)-β-L- arabinan through direct interaction with thrombin
topic_facet	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
description	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.
author	Fernández, Paula Virginia Quintana, Irene Luisa Cerezo, Alberto Saúl Estevez, Jose Manuel Ciancia, Marina
author_facet	Fernández, Paula Virginia Quintana, Irene Luisa Cerezo, Alberto Saúl Estevez, Jose Manuel Ciancia, Marina
author_sort	Fernández, Paula Virginia
title	Anticoagulant activity of a unique sulfated pyranosic (1→3)-β-L- arabinan through direct interaction with thrombin
title_short	Anticoagulant activity of a unique sulfated pyranosic (1→3)-β-L- arabinan through direct interaction with thrombin
title_full	Anticoagulant activity of a unique sulfated pyranosic (1→3)-β-L- arabinan through direct interaction with thrombin
title_fullStr	Anticoagulant activity of a unique sulfated pyranosic (1→3)-β-L- arabinan through direct interaction with thrombin
title_full_unstemmed	Anticoagulant activity of a unique sulfated pyranosic (1→3)-β-L- arabinan through direct interaction with thrombin
title_sort	anticoagulant activity of a unique sulfated pyranosic (1→3)-β-l- arabinan through direct interaction with thrombin
publishDate	2013
url	https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00219258_v288_n1_p223_Fernandez http://hdl.handle.net/20.500.12110/paper_00219258_v288_n1_p223_Fernandez
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Anticoagulant activity of a unique sulfated pyranosic (1→3)-β-L- arabinan through direct interaction with thrombin

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