title: Evaluation of biomolecular interactions of sulfated polysaccharide isolated from Grateloupia filicina on blood coagulation factors.
abstract: An edible marine red alga, Grateloupia filicina, collected from Jeju Island of Korea was hydrolyzed by cheap food-grade carbohydrases (Viscozyme, Celluclast, AMG, Termamyl, and Ultraflo) to investigate its anticoagulant activity. Among the tested enzymatic extracts of G. filicina, a Termamyl extract showed the highest anticoagulant activity. Anion-exchange chromatography on DEAE-cellulose and gelpermeation chromatography on Sepharose-4B were used to purify the active polysaccharide from the crude polysaccharide fraction of G. filicina. The purified sulfated polysaccharide (0.42 sulfate/total sugar) showed approximately 1,357 kDa molecular mass and was comprised mainly of galactose (98%) and 1-2% of glucose. The sample showed potential anticoagulant activity on activated partial thromboplastin time (APTT) and thrombin time (TT) assays. The purified G. filicina anticoagulant (GFA) inhibited the coagulation factor X (92%), factor II (82%), and factor VII (68%) of the coagulation cascade, and the molecular interaction (protein-polysaccharide) was highly enhanced in the presence of ATIII (antithrombin III). The dissociation constant of polysaccharide towards serine proteins decreased in the order of FXa (58.9 nM) >FIIa (74.6 nM) >FVII (109.3 nM). The low/less cytotoxicity of the polysaccharide benefits its use in the pharmaceutical industry; however, further studies that would help us to elucidate the mechanism of its activity are needed.
authors.affiliation: Faculty of Applied Marine Science, Cheju National University, Jeju 690-756, Korea.
authors.forename: Yasantha
authors.initials: Y
authors.lastname: Athukorala
authors.forename: Won-Kyo
authors.initials: WK
authors.lastname: Jung
authors.forename: Pyo-Jam
authors.initials: PJ
authors.lastname: Park
authors.forename: Young-Jae
authors.initials: YJ
authors.lastname: Lee
authors.forename: Se-Kwon
authors.initials: SK
authors.lastname: Kim
authors.forename: Thava
authors.initials: T
authors.lastname: Vasanthan
authors.forename: Hong-Kyoon
authors.initials: HK
authors.lastname: No
authors.forename: You-Jin
authors.initials: YJ
authors.lastname: Jeon
mesh_terms: D000818:Animals; D000925:Anticoagulants / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism* / Q000494:pharmacology; D001779:Blood Coagulation Factors / Q000037:antagonists & inhibitors* / Q000378:metabolism*; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D006224:Cricetinae; D006493:Heparin / Q000378:metabolism / Q000494:pharmacology; D006801:Humans; D007700:Kinetics; D007723:Korea; D011134:Polysaccharides / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism* / Q000494:pharmacology; D000461:Rhodophyta / Q000737:chemistry / Q000378:metabolism*
chemical_list: D000925:Anticoagulants; D001779:Blood Coagulation Factors; D011134:Polysaccharides; D006493:Heparin