title: Novel thrombolytic protease from edible and medicinal plant Aster yomena (Kitam.) Honda with anticoagulant activity: purification and partial characterization.
abstract: A thrombolytic protease named kitamase possessing anticoagulant property was purified from edible and medicinal plant Aster yomena (Kitam.) Honda. Kitamase showed a molecular weight of 50 kDa by SDS-PAGE and displayed a strong fibrin zymogram lysis band corresponding to the similar molecular mass. The enzyme was active at high temperatures (50°C). The fibrinolytic activity of kitamase was strongly inhibited by EDTA, EGTA, TPCK and PMSF, inhibited by Zn(2+). The Km and Vmax values for substrate S-2251 were determined as 4.31 mM and 23.81 mM/mg respectively. It dissolved fibrin clot directly and specifically cleaved the α, Aα and γ-γ chains of fibrin and fibrinogen. In addition, kitamase delayed the coagulation time and increased activated partial thromboplastin time and prothrombin time. Kitamase exerted a significant protective effect against collagen and epinephrine induced pulmonary thromboembolism in mice. These results suggest that kitamase may have the property of metallo-protease like enzyme, novel fibrino(geno)lytic enzyme and a potential to be a therapeutic agent for thrombosis.
authors.affiliation: Department of Life Science & BK21-Plus Research Team for Bioactive Control Technology, Chosun University, Gwangju 501-759, Republic of Korea.
authors.forename: Jun-Hui
authors.initials: JH
authors.lastname: Choi
authors.affiliation: Department of Life Science & BK21-Plus Research Team for Bioactive Control Technology, Chosun University, Gwangju 501-759, Republic of Korea.
authors.forename: Dae-Won
authors.initials: DW
authors.lastname: Kim
authors.affiliation: Department of Life Science & BK21-Plus Research Team for Bioactive Control Technology, Chosun University, Gwangju 501-759, Republic of Korea.
authors.forename: Se-Eun
authors.initials: SE
authors.lastname: Park
authors.affiliation: Jangheung Research Institute for Mushroom Industry, Jangheung 529-851, Republic of Korea.
authors.forename: Bong-Suk
authors.initials: BS
authors.lastname: Choi
authors.affiliation: Department of Life Science & BK21-Plus Research Team for Bioactive Control Technology, Chosun University, Gwangju 501-759, Republic of Korea; Central Department of Zoology, Tribhuvan University, Kirtipur, Kathmandu, Nepal.
authors.forename: Kumar
authors.initials: K
authors.lastname: Sapkota
authors.affiliation: Department of Alternative Medicine, Gwangju University, Gwangju 503-703, Republic of Korea.
authors.forename: Seung
authors.initials: S
authors.lastname: Kim
authors.affiliation: Department of Life Science & BK21-Plus Research Team for Bioactive Control Technology, Chosun University, Gwangju 501-759, Republic of Korea. Electronic address: sjbkim@chosun.ac.kr.
authors.forename: Sung-Jun
authors.initials: SJ
authors.lastname: Kim
mesh_terms: D000818:Animals; D031173:Aster Plant / Q000737:chemistry* / Q000201:enzymology; D001780:Blood Coagulation Tests; D002413:Cations, Divalent; D003094:Collagen; D004492:Edetic Acid / Q000737:chemistry; D004533:Egtazic Acid / Q000737:chemistry; D010450:Endopeptidases / Q000302:isolation & purification* / Q000378:metabolism / Q000494:pharmacology; D005337:Fibrin / Q000737:chemistry / Q000378:metabolism; D005340:Fibrinogen / Q000737:chemistry / Q000378:metabolism; D005342:Fibrinolysis / Q000187:drug effects; D005343:Fibrinolytic Agents / Q000302:isolation & purification* / Q000378:metabolism / Q000494:pharmacology; D006358:Hot Temperature; D007700:Kinetics; D008297:Male; D051379:Mice; D008813:Mice, Inbred ICR; D008970:Molecular Weight; D018515:Plant Leaves / Q000737:chemistry; D010940:Plant Proteins / Q000302:isolation & purification* / Q000378:metabolism / Q000494:pharmacology; D010946:Plants, Medicinal; D011655:Pulmonary Embolism / Q000097:blood / Q000139:chemically induced / Q000188:drug therapy*; D015032:Zinc / Q000737:chemistry
chemical_list: D002413:Cations, Divalent; D005343:Fibrinolytic Agents; D010940:Plant Proteins; D004533:Egtazic Acid; D005337:Fibrin; D005340:Fibrinogen; D003094:Collagen; D004492:Edetic Acid; D010450:Endopeptidases; D015032:Zinc
keywords: Anticoagulant; Antithrombotic; Aster yomena (Kitam.) Honda; Fibrinolysis; Medicinal plant