Isolation and partial purification of anticoagulant fractions from the venom of the Iranian snake Echis carinatus

• Autorzy: Babaie M. , Zolfagharian H. , Salmanizadeh H. , Mirakabadi A. Z. , Alizadeh H.
• Języki publikacji: EN

Abstrakty

EN

 Many snake venoms comprise different factors, which can either promote or inhibit the blood coagulation pathway. Coagulation disorders and hemorrhage belong to the most prominent features of bites of the many vipers. A number of these factors interact with components of the human blood coagulation. This study is focused on the effect of Echis carinatus snake venom on blood coagulation pathway. Anticoagulant factors were purified from the Iranian Echis carinatus venom by two steps: gel filtration (Sephadex G-75) and ion-exchange (DEAE-Sephadex) chromatography, in order to study the anticoagulant effect of crude venom and their fractions. The prothrombin time was estimated on human plasma for each fraction. Our results showed that protrombin time value was increase from 13.4 s to 170 s for F2C and to 280 s for F2D. Our study showed that these fractions of the venom delay the prothrombine time and thus can be considered as anticoagulant factors. They were shown to exhibit proteolytic activity. The molecular weights of these anticoagulants (F2C, F2D) were estimated by SDS/PAGE electrophoresis. F2C comprises two protein bands with molecular weights of 50 and 79 kDa and F2D a single band with a molecular weight of 42 kDa.

• Wydawca: -
• Czasopismo: Acta Biochimica Polonica
• Rocznik: 2013
• Tom: 60
• Numer: 1
• Opis fizyczny: p.17-20,fig.,ref.

Twórcy

autor

Babaie M.

• Department of Biology, Science & Research Branch, Islamic Azad University, Tehran, Iran

autor

Zolfagharian H.

• Department of Venomous Animals and Antivenom Production, Razi Vaccine and Serum Research Institute, Alborz, Karaj, Iran

autor

Salmanizadeh H.

• Department of Biology, Science & Research Branch, Islamic Azad University, Tehran, Iran

autor

Mirakabadi A. Z.

• Department of Venomous Animals and Antivenom Production, Razi Vaccine and Serum Research Institute, Alborz, Karaj, Iran

autor

Alizadeh H.

• Department of Biology, Science & Research Branch, Islamic Azad University, Tehran, Iran

Bibliografia

• Daisuke Y, Fujio S, Takashi M (1996) Isolation and characterization of carinactivase, a novel prothrombin activator in Echis carinatus venom with a unique catalytic mechanism. J Biol Chem 271: 5200-5207. 
• Higuchi DA, Barbosa CM, Bincoletto C, Chagas JR, Magalhaes A, Richardson M (2007) Purification and partial characterization of two phospholipases A2 from Bothrops leucurus (white-tailed jararaca) snake venom. Biochimie 89: 319-328. 
• Janos P, Edith MM, Manuel CP, Timothy NCW, Mansoor SAS, Kenneth JC (1997) Amino acid sequence of the α subunit and computer modelling of the α and β subunits of echicetin from the venom of Echis carinatus (saw-scaled viper). Biochem J 323: 533-537. 
• Julian W (2005) Snake venoms and coagulopathy. Toxicon 45: 951-967. 
• Kini RM (2005) Serine proteases affecting blood coagulation and fibrinolysis from snake venoms. Pathophysiol Haemost Thromb 34: 200-204. 
• Kini RM (2005) Structure-function relationships and mechanism of anticoagulant phospholipase A2 enzymes from snake venoms. Toxicon 45: 1147-1153. 
• Kini RM (2006) Anticoagulant proteins from snake venoms: structure, function and mechanism. Biochem J 397: 377-387. 
• Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680-685. 
• Loubna H, Cesare G, Silvia C, Stefano R, Fatima LD (2010) Purification and characterization of a fibrinogenolytic and hemorrhagic metalloproteinase isolated from Vipera lebetina venom. Biochimie 92: 797-784. 
• Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193: 265-275. 
• Markland FS (1998) Snake venoms and the hemostatic system. Toxicon 36: 1749-1800. 
• Morita T (2005) Structures and functions of snake venom CLPs (C-type lectin-like proteins) with anticoagulant procoagulant and plateletmodulating activities. Toxicon 45: 1099-1105. 
• Oyama E, Takahashi H (2003) Purification and characterization of a thrombin-like enzyme, elegaxobin II, with lys-bradykinin releasing activity from the venom of Trimeresurus elegans (Sakishima-Habu). Toxicon 41: 559-568. 
• Peichoto ME, Teibler P, Mackessy SP, Leiva L, Acosta O, Gonçalves LR (2007) Purification and characterization of patagonfibrase, a metalloproteinase showing α-fibrinogenolytic and hemorrhagic activities, from Philodryas patagoniensis snake venom. Biochim Biophys Acta 1770: 810-817. 
• Qing-qiu H, Mai-kun, Li-wen N (1999) Purification and characterization of two fibrinogen-clotting enzymes from five-pace snake (Agkistrodon acutus) venom. Toxicon 37: 999-1013. 
• Rizzo F, Papasouliotis K, Crawford E, Dodkin S, Cue S (2008) Measurement of prothrombin time (PT) and activated partial thromboplastin time (APTT) on canine citrated plasma samples following different storage conditions. Res Vet Sci 85: 166-170. 
• Wang Y, Cui G, Zhao M, Yang J, Wang C, Giese RW (2008) Bioassay-directed purification of an acidic phospholipase A2 from Agkistrodon halys pallas venom. Toxicon 51: 1131-1139. 
• Wei XL, Wei JF, Li T, Qiao LY, Liu YL, Huang T (2007) Purification, characterization and potent lung lesion activity of an L-amino acid oxidase from Agkistrodon blomhoffii ussurensis snake venom. Toxicon 50: 1126-1139. 

Uwagi

rekord w opracowaniu