Isolation and characterization of a fibrinogen-clotting enzyme from venom of the snake, Lachesis muta muta(Peruvian bushmaster)

Yarlequé, Armando; Campos, Sylvia Maria Nicolau; Escobar, Enrique; Lazo, Fanny; Sánchez, Norma Silvia; Hyslop, Stephen; Na, Marsh; Butterworth, Peter J.; Price, Robert G.

doi:10.1016/0041-0101(89)90027-5

Cited by 29 publications

(13 citation statements)

References 14 publications

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“…TLBbar showed of 0.35 nmoles/min on BApNA substrate (Figure 4(a)), with Michaelis-Menten kinetic behavior (Figure 4(c)) and max and of 0.42 nmol/min and 0.433 mM, respectively (Figure 4(d)); these values are similar to other thrombin-like snake venom, as TLE-1 and TLE-2 isolated from B. atrox ( 0.33 mM and 0.46 mM, resp.) [37] and thrombin-like from Lachesis muta muta ( of 0.075 mM and max of 0.36 nmol/min) [38]. These results suggested that the catalytic mechanism of this enzyme is probably involved in an active site organization similar to that of trypsin.…”

Section: Discussionmentioning

confidence: 55%

Biochemical and Pharmacological Characterization of TLBbar, a New Serine Protease with Coagulant Activity fromBothrops barnettiSnake Venom

et al. 2013

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A thrombin-like enzyme named TLBbar was isolated from Bothrops barnetti snake venom and its biochemical and pharmacological characteristics were determined. TLBbar was purified using size exclusion chromatography and reverse phase HPLC, showing molecular mass of 28750.7 Da determined by mass spectrometry. TLBbar serine protease is basic (pI 7.4) and its structure shows similarity with other serine proteases of snake venom. Optimal proteolytic activity was at 37 ∘ C and pH 8; this activity was strongly inhibited by PMSF and Leupeptin, however; heparin, and soybean trypsin inhibitor (SBT-I) were ineffective. Kinetic studies on BApNA chromogenic substrate have revealed that TLBbar presents a Michaelis-Menten kinetics, with values of and max of 0.433 mM and 0.42 nmol/min, respectively. TLBbar showed high clotting activity upon bovine and human plasma, presenting IC of 125 and minimum dose coagulant (MDC) of 2.23 g/ L. TLBbar cleavages the A chain of bovine fibrinogen, with maximal efficiency at 30-40 ∘ C in the presence of calcium after two hours incubation; this fibronogenolityc activity was inhibited by PMSF and Leupeptin, confirming its classification in the group of serine proteases. In addition, TLBbar is capable of aggregating platelets in the same way that thrombin in concentrations of 2.5 g/ L.

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Section: Discussionmentioning

confidence: 55%

Biochemical and Pharmacological Characterization of TLBbar, a New Serine Protease with Coagulant Activity fromBothrops barnettiSnake Venom

et al. 2013

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“…Se determinó por el método descrito por Yarlequé et al (1989). La mezcla de reacción contenía 0,2 mL de plasma humano y 0,1 mL de la enzima, se incubaron a 37 ºC por 5 min para luego agregarles 0,1 mL de CaCl 2 25 mM midiéndose el tiempo de recalcificación.…”

Section: B) Actividad Hemolítica Indirecta En Placaunclassified

Caracterización biológica y acción de inhibidores de una fosfolipasa A2 del veneno de Lachesis muta

et al. 2011

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“…Cabe mencionar que las proteínas estudia das de venenos de serpientes del Perú se han tralado exclusivamente de proteínas con acti vidad enzirnática (Yarlequé et aL, 1989;Pa ntigo o et al, 1996). EI,l particular algunas de las proteínas que s han aislado y caracte~ rizad en el veneno de B bra::ili, son las e nzimas similar a trombina (Limán, 1996), fos l'ol ipasa (Zeballos el al.…”

Section: Introductionunclassified

Aislamiento y caracterización de una miotoxina del veneno de la serpiente Bothrops brazili Hoge, 1953 (Ophidia: Viperidae)

2001

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RESUMENSe ha purificado una miotoxina del veneno de la serpiente BOl/1ropS brazifi, empleando un solo pas cromatográfico de intercambio jónico sobre CM-Sephadex C-50 con buffer acetato de amonio 0,05 M pH 7. La pureza de la proteina fue evaluada por PAGE con y sin SOS , inm unodifusión inmunoeleclroforesis . La proteína es de naturaleza basica y contiene 15,6% de Lys+Arg ; además, n esta glicosilada , carece de actividad enzimatica , y por el método de Lowry se ha calculado que ell cons tituye el 25% de la proteína tolal del veneno .Por PAG E-SOS y cromatografía de fi ltración, se ha determinado que la miotoxina tiene un pes molecula r de 30 KOa , y está formada por 2 cadenas polipeplidicas de 15 KOa cada una.La inocu lación de la miotoxina en el músculo gastrocnemi us de ratones albinos , produce una seve ra necrosis del tejido .La miotoxina no tiene actividad hemol itica ni anticoagu la nte; sin em bargo , si produce edema , se h calculado una OEM de 32,6 ¡.¡g de proteína .Palabras clave : miotoxina, BOlhrops brazifi, veneno de serpiente, mionecrosis ABSTRACT A myotoxin from the venom of the snake Bothrops brazifi has been purified by io n-excha ng chro matography on CM-Sephadex C-50 with 0,05 M ammo nium acelate buffer pH 7. The homogene il was evaluated by PAGE wilh and withoul SOS, immunodi ffusion and immunoeleclrophoresis. Th myotoxin is a basic protein with 15,6% of Lys+Arg; il is not a glicoprotein , has not enzymatic activity, an corresponds to 25% of the whole venom protein.The molecularweight ofthe myotoxin was determined by PAGE-SOS and gel filtraUon chromatography The myotoxin has 30 KDa of molecular weight and two polypeptide chains of 15 KOa each .Myotoxin produces a severe necrosis on the gastrocnemius muscle of white mice.Th e myotoxin does not have hemolytic nor anticoagulant activity. However, produces edema with OEM of 32,6 [.1g of protein .

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Isolation and characterization of a fibrinogen-clotting enzyme from venom of the snake, Lachesis muta muta(Peruvian bushmaster)

Cited by 29 publications

References 14 publications

Biochemical and Pharmacological Characterization of TLBbar, a New Serine Protease with Coagulant Activity fromBothrops barnettiSnake Venom

Biochemical and Pharmacological Characterization of TLBbar, a New Serine Protease with Coagulant Activity fromBothrops barnettiSnake Venom

Caracterización biológica y acción de inhibidores de una fosfolipasa A2 del veneno de Lachesis muta

Aislamiento y caracterización de una miotoxina del veneno de la serpiente Bothrops brazili Hoge, 1953 (Ophidia: Viperidae)

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