Partial purification of the plasma substrate for the bradykinin-releasing enzyme from the venom of Bothrops jararaca

Brasil 1Bothrops jararaca venom (BJV) caused a fall in the carotid artery blood pressure of the anaesthetized snake. This effect was tachyphylactic and was potentiated by captopril, a kininase II inhibitor; it was partially antagonized by promethazine plus cimetidine and was not affected by atropine. 2 Similar hypotensive effects were obtained by administration of trypsin or a partially purified BJV kininogenase to the snake. 3 Incubation of Bothrops jararaca plasma (BJP) with trypsin released a substance (or substances) that produced hypotension in the snake but not in the rat; this hypotensive effect was also potentiated by captopril. 4 The trypsinised plasma contracted Bothrops jararaca isolated uterus, a pharmacological preparation weakly sensitive to bradykinin. Trypsinised plasma was inactive on pigeon oviduct and rat uterus and displayed a weak action on the guinea-pig ileum. Similar effects were observed with incubates of a fraction of BJP, containing globulins, with a partially purified BJV kininogenase. 5 Like mammalian kinins, the substance(s) was(were) dialysable, thermostable in acid but not in alkaline pH, and inactivated by chymotrypsin but not by trypsin. Its(their) inactivation by BJP or BJP kininase II was inhibited by captopril. 6 These findings strongly suggest that, besides releasing histamine, BJV or trypsin release a kininlike substance (or substances) from the snake plasma. 7 Since BJV and other kininogenases active on mammalian plasmas were shown to be unable to release kinins from BJP, in experiments conducted on pharmacological preparations suitable for the assay of mammalian kinins, these data also suggest that the snake Bothrops jararaca, like birds, may have developed its own kallikrein-kinin system.

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“…The rat uterus was prepared according to Gaddum & Hameed (1954) and the guinea-pig ileum as described by Henriques et al (1962).…”

Section: Isolated Smooth Muscle Preparationsmentioning

confidence: 99%

Kallikrein‐kinin system in the plasma of the snake Bothrops jararaca

Abdalla

Hiraichi

Picarelli

et al. 1989

British J Pharmacology

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“…Crotalidae venoms release kinins [3]; their .effects were thus also compared to those of trypsin, known to release bradykinin, and to those of a 'bradykinin releasing enzyme' (BRE) purified from Bothrops jararaca venom [4].…”

Section: Introductionmentioning

confidence: 99%

Haemorrhagic and permeability increasing effects of ‘Bothrops jararaca’ and other crotalidae venoms as related to amine or kin in release

1974

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The venom 'Bothrops jararaca' anda high molecular weight fraction obtained from it induced vasculotoxic effects when applied on the exposed surface of the lung of the dog and increased the vascular permeability when injected into the rat skin. These effects were compared to those evoked by other venoms and potential mediators. The increase in vessel permeability due to 'B. jararaca' venom was feebly affected by antiserotonin and antihistamine agents and was presumably due to kinin release. The effect on the lung surface was related to the coagulant and necrogenic properties of the venom and was suppressed by EDTA, promethazine and specific antiserum. A kinin releasing enzyme extracted from the venom increased the vascular permeability but had no effect on the lung. Venoms from 'Crotalus adamanteus' and from 'Crotalus durissus terrificus' also increased the vascular permeability, but were inhibited by antihistamine/antiserotonin agents; this blockade uncovered a marked skin hemorrhagic effect due to 'Crotalus adamanteus'. Despite this activity, no vasculotoxic effect was found when 'Crotalus adamanteus' venom was applied to the lung. Effects of the tested venoms and enzymes are highly specific for either vascular bed, depending upon the availability on the challenged sites of different tissue components and on the specific properties of the venoms.

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“…Purified horse bradykininogen was said to be free from kinin-forming enzyme and assumed to be free from kininase. No supporting data were published however (Henriques, Picarelli & Ferraz de Oliveira, 1962). Purified bovine bradykininogen was free from kinin-forming enzyme but contained kininase (Habermann & Rosenbusch, 1962).…”

Section: Development Of a Suitable Purification Proceduresmentioning

confidence: 98%

“…The most hazardous step is probably the solution of the freeze-dried protein and buffer salts prior to gel filtration on G-200 Sephadex. The acid treatment of horse plasma (Henriques et al, 1962) may cause irreversible changes in the tertiary structure of bradykininogen. The heat coagulation used by Greenbaum & Hosoda (1963) probably denatures bradykininogen.…”

Section: Development Of a Suitable Purification Proceduresmentioning

confidence: 99%

“…Henriques, Picarelli & Ferraz de Oliveira (1962) treated horse plasma with acid, and fractionated by precipitation with ammonium sulphate and chromatography on DEAE-cellulose. The product contained 34% of the bradykininogen present in the acid-treated plasma, and 1 mg of the protein gave the equivalent of 3.14 Fkg bradykinin with trypsin and 5.0 ,ug with snake venom.…”

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confidence: 99%

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The Purification of a Kininogen From Human Plasma

Brocklehurst¹,

Mawer²

1966

British Journal of Pharmacology and Chemotherapy

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Several groups of workers have described methods for the purification of plasma bradykininogen. Henriques, Picarelli & Ferraz de Oliveira (1962) treated horse plasma with acid, and fractionated by precipitation with ammonium sulphate and chromatography on DEAE-cellulose. The product contained 34% of the bradykininogen present in the acid-treated plasma, and 1 mg of the protein gave the equivalent of 3.14 Fkg bradykinin with trypsin and 5.0 ,ug with snake venom.Habermann & Rosenbusch (1962) purified bovine plasma bradykininogen by Cohn method 6 and chromatography on calcium hydroxylapatite. The product had a specific activity of 10 to 20 Mg bradykinin per mg protein.Greenbaum & Hosoda (1963) purified bovine plasma bradykininogen by ammonium sulphate precipitation, heat coagulation and iso-electric precipitation. The product contained less than 3% of the bradykininogen present in the total plasma globulin and had a specific activity of 0.8 to 1.0 ,ug bradykinin per mg protein.Webster & Pierce (1963) purified bradykininogen from stored human plasma by chromatography on DEAE-cellulose and calcium hydroxylapatite. The product contained 15% of the bradykininogen present in stored plasma but further losses occurred during dialysis, filtration and freeze-drying.The purpose of the present communication is to report a method for the purification of human plasma bradykininogen which has advantages over the earlier methods.METHODS The method consists of three main steps.1. Venous blood was collected and the plasma rapidly separated from the cells. The plasma proteins were equilibrated with phosphate buffer on a column of G-50 Sephadex in preparation for ion exchange chromatography.2. The part of the eluate which contained the bradykininogen was applied to a column of DEAE-Sephadex. A bradykininogen enriched sample (fraction C) was eluted by stepwise increases in salt concentration. This fraction was suitable for many pharmacological purposes without further treatment.3. After concentration by freeze drying the bradykininogen of fraction C was purified further by gel filtration on G-200 Sephadex.

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Partial purification of the plasma substrate for the bradykinin-releasing enzyme from the venom of Bothrops jararaca

Cited by 35 publications

References 10 publications

Kallikrein‐kinin system in the plasma of the snake Bothrops jararaca

Kallikrein‐kinin system in the plasma of the snake Bothrops jararaca

Haemorrhagic and permeability increasing effects of ‘Bothrops jararaca’ and other crotalidae venoms as related to amine or kin in release

The Purification of a Kininogen From Human Plasma

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