The complete amino acid sequence of a thrombin‐like enzyme/gyroxin analogue from venom of the bushmaster snake (<i>Lachesis muta muta</i>)

Magalhães, Ana C.; Fonseca, B.C.B.; Diniz, Carlos R.; Gilroy, John; Richardson, Michael

doi:10.1016/0014-5793(93)80205-9

Cited by 53 publications

(14 citation statements)

References 25 publications

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“…The homology study (Figure 4) showed that Rhombeobin shared a high degree of sequence identity with SVSP with thrombin-like activity [12, 13, 15–18, 28, 35] especially with LM-TL ( Lachesis muta muta ). Detailed analysis showed small but important differences between Rhombeobin and LM-TL.…”

Section: Discussionmentioning

confidence: 99%

Rapid Purification and Procoagulant and Platelet Aggregating Activities of Rhombeobin: A Thrombin-Like/Gyroxin-Like Enzyme fromLachesis muta rhombeataSnake Venom

Torres-Huaco

Werneck²,

Vicente³

et al. 2013

BioMed Research International

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We report a rapid purification method using one-step chromatography of SVSP Rhombeobin (LMR-47) from Lachesis muta rhombeata venom and its procoagulant activities and effects on platelet aggregation. The venom was fractionated by a single chromatographic step in RP-HPLC on a C8 Discovery BIO Wide Pore, showing high degree of molecular homogeneity with molecular mass of 47035.49 Da. Rhombeobin showed amidolytic activity upon BAρNA, with a broad optimum pH (7–10) and was stable in solution up to 60°C. The amidolytic activity was inhibited by serine proteinase inhibitors and reducing agents, but not chelating agents. Rhombeobin showed high coagulant activity on mice plasma and bovine fibrinogen. The deduced amino acid sequence of Rhombeobin showed homology with other SVSPs, especially with LM-TL (L. m. muta) and Gyroxin (C. d. terrificus). Rhombeobin acts, in vitro, as a strong procoagulant enzyme on mice citrated plasma, shortening the APTT and PT tests in adose-dependent manner. The protein showed, “ex vivo”, a strong defibrinogenating effect with 1 µg/animal. Lower doses activated the intrinsic and extrinsic coagulation pathways and impaired the platelet aggregation induced by ADP. Thus, this is the first report of a venom component that produces a venom-induced consumptive coagulopathy (VICC).

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Section: Discussionmentioning

confidence: 99%

Rapid Purification and Procoagulant and Platelet Aggregating Activities of Rhombeobin: A Thrombin-Like/Gyroxin-Like Enzyme fromLachesis muta rhombeataSnake Venom

Torres-Huaco

Werneck²,

Vicente³

et al. 2013

BioMed Research International

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“…Evidence has accumulated in our lab that has allowed us to safely presume that the 45-47 kDa species isolated from L. m. rhombeata venom using chromatography on Am-PABA resin is virtually identical to the ortologue from L. m. muta venom that had it primary structure determined [36,45]. In fact, the very existence of these subspecies seems not to be a consensus among herpetologists [46].…”

Section: Computer-aided Evaluation Of the Interaction Between Lmm-tlementioning

confidence: 99%

“…For the ligand representing Table 2 Comparison of inhibition constants (K i ) of S1-directed inhibitors of thrombin-like serine proteases (TLSPs) from Bothrops jararacussu (BJ-48) and Lachesis muta muta (Lmm-TLE) venoms S1-directed inhibitor b Constants at 25 • C using BapNA as substrate [45]. Values are shown as means ± standard deviations.…”

Section: Computer-aided Evaluation Of the Interaction Between Lmm-tlementioning

confidence: 99%

Biochemical and molecular modeling analysis of the ability of two p-aminobenzamidine-based sorbents to selectively purify serine proteases (fibrinogenases) from snake venoms

De-Simone¹,

Corrêa-Netto²,

Antunes³

et al. 2005

Journal of Chromatography B

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“…The enzyme is a single polypeptide chain glycoprotein with Mr of 33 kDa, and belongs to the trypsin family of serine proteinases. The partial amino acid sequence (77%) of the enzyme reveals that it shares structural homology with other serine proteinases isolated from the same source including the plasminogen activator (LV-PA) (15) and the clotting enzyme (16) as well as with other serine proteinases from snake venoms and mammalian kallikrein (8,17).…”

Section: Introductionmentioning

confidence: 99%

Biochemical Properties of a Bushmaster Snake Venom Serine Proteinase (LV-Ka), and Its Kinin Releasing Activity Evaluated in Rat Mesenteric Arterial Rings

et al. 2004

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Abstract. A serine proteinase with kallikrein-like activity (LV-Ka) has been purified to homogeneity from bushmaster snake (Lachesis muta muta) venom. Physicochemical studies indicated that LV-Ka is a single chain glycoprotein with a molecular mass (Mr) of 33 kDa under reducing conditions which was reduced to 28 kDa after treatment with N-Glycosidase F (PNGase F). LV-Ka can be bounded and neutralized by serum a 2 -macroglobulin (a 2 -M), a prevalent mammalian protease inhibitor that is capable of forming a macromolecular complex with LV-Ka (Mr >180 kDa). Cleavage of a 2 -M by the enzyme resulted in the formation of 90-kDa fragments. The proteolytic activity of LV-Ka against dimethylcasein could be inhibited by a 2 -M, and the binding ratio of the inhibitor:enzyme complex was found to be 1:1. The Michaelis constant, K m , and catalytic rate constant, kcat, of LV-Ka on four selective chromogenic substrates were obtained from Lineweaver-Burk plots. LV-Ka exhibits substrate specificities not only for the glandular kallikrein H-D-Val-Leu-Arg-pNA (S-2266) but also for the plasmin substrates S-2251 and Tos-Gly-Pro-Lys-pNA. Bovine kininogen incubated with LV-Ka generated a polypeptide that dose dependently contracted mesenteric arterial rings from spontaneously hypertensive rats (SHR) in a similar way as bradykinin (BK) does. As it happens with BK, LV-Ka generated polypeptide was inhibited by HOE-140, a bradykinin B 2 -receptor antagonist and by indomethacin, a cyclo-oxygenase inhibitor. These results strongly suggest that the polypeptide generated by LV-Ka by cleavage of bovine kininogen is bradykinin. In addition, our studies may help to understand the mechanism of action involved in hypotension produced by envenomation of bushmaster snake.

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The complete amino acid sequence of a thrombin‐like enzyme/gyroxin analogue from venom of the bushmaster snake (Lachesis muta muta)

Cited by 53 publications

References 25 publications

Rapid Purification and Procoagulant and Platelet Aggregating Activities of Rhombeobin: A Thrombin-Like/Gyroxin-Like Enzyme fromLachesis muta rhombeataSnake Venom

Rapid Purification and Procoagulant and Platelet Aggregating Activities of Rhombeobin: A Thrombin-Like/Gyroxin-Like Enzyme fromLachesis muta rhombeataSnake Venom

Biochemical and molecular modeling analysis of the ability of two p-aminobenzamidine-based sorbents to selectively purify serine proteases (fibrinogenases) from snake venoms

Biochemical Properties of a Bushmaster Snake Venom Serine Proteinase (LV-Ka), and Its Kinin Releasing Activity Evaluated in Rat Mesenteric Arterial Rings

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