title: Highly sulfated dermatan sulfate from the skin of the ray Raja montagui: anticoagulant activity and mechanism of action.
abstract: The dermatan sulfate (DS) isolated from the ray skin Raja montagui was identified and characterized. Its average molecular weight (Mw) and sulfate content were 39 kDa and 25% w/w, respectively. This DS prolonged thrombin time and activated partial thromboplastin time and inhibited the thrombin generation in a concentration-dependent manner whereas it had no effect on the anti-Xa assay and on platelet function. Data from the anti-IIa assay allowed the assessment of the specific anticoagulant activity which was 40 units/mg. The kinetics of the thrombin inhibition by heparin cofactor II (HCII) has been studied as a function of DS concentration according to a kinetic model in which the polysaccharide binds quickly to the inhibitor and forms a complex more reactive than the free inhibitor towards thrombin. This DS accelerated thrombin inhibition exclusively by HCII. The dissociation constant of the DS-HCII complex, K(DSHCII), and the rate constant of the thrombin inhibition by this complex, k, were (2.93+/-0.25)x10(-6)M and (2.2+/-0.35)x10(9)M(-1)min(-1), respectively. Our findings indicated that the major polysaccharide in the skin of the ray Raja montagui was a DS endowed with a high anticoagulant effect mediated by HCII and which may constitute an anticoagulant drug of interest in anticoagulant therapy.
authors.affiliation: Laboratoire de Pharmacologie 04/UR/01-09, Faculté de Médecine, 5000 Monastir, Tunisia.
authors.forename: Mohamed
authors.initials: M
authors.lastname: Ben Mansour
authors.forename: Manel
authors.initials: M
authors.lastname: Dhahri
authors.forename: Mohsen
authors.initials: M
authors.lastname: Hassine
authors.forename: Nadine
authors.initials: N
authors.lastname: Ajzenberg
authors.forename: Laurence
authors.initials: L
authors.lastname: Venisse
authors.forename: Véronique
authors.initials: V
authors.lastname: Ollivier
authors.forename: Frédéric
authors.initials: F
authors.lastname: Chaubet
authors.forename: Martine
authors.initials: M
authors.lastname: Jandrot-Perrus
authors.forename: Raoui M
authors.initials: RM
authors.lastname: Maaroufi
mesh_terms: D000818:Animals; D000925:Anticoagulants / Q000737:chemistry* / Q000302:isolation & purification / Q000494:pharmacology*; D003871:Dermatan Sulfate / Q000737:chemistry* / Q000302:isolation & purification / Q000494:pharmacology*; D065427:Factor Xa Inhibitors; D015844:Heparin Cofactor II / Q000494:pharmacology; D015539:Platelet Activation / Q000187:drug effects; D010974:Platelet Aggregation / Q000187:drug effects; D011516:Prothrombin / Q000037:antagonists & inhibitors; D012860:Skates, Fish; D012867:Skin / Q000737:chemistry; D013431:Sulfates / Q000032:analysis; D013917:Thrombin / Q000378:metabolism
chemical_list: D000925:Anticoagulants; D065427:Factor Xa Inhibitors; D013431:Sulfates; D003871:Dermatan Sulfate; D015844:Heparin Cofactor II; D011516:Prothrombin; C025337:Factor IIa; D013917:Thrombin