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      Comparison of proteins, lethality and immunogenic compounds of Androctonus crassicauda (Olivier, 1807) (Scorpiones: Buthidae) venom obtained by different methods

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          Abstract

          Scorpions are venomous arthropods of the class Arachnida and are considered relatives of spiders, ticks and mites. There are approximately 1,500 species of scorpions worldwide, which are characterized by an elongated body and a segmented tail that ends in a venomous stinger. No specific treatment is available for scorpion envenomation, except for the use of antivenom. The current study aimed at comparing protein content and lethality of Androctonus crassicauda venom extracted by two different methods (electric stimulation and maceration of telsons). The LD50 calculated by probit analysis was 1.1mg/kg for venom obtained by electric stimulation and 39.19mg/kg for venom obtained by maceration of telsons. In the electrophoretic analysis, protein bands of the venom sample obtained by electric stimulation were between 12 and 53kDa (total: five bands), and those of venom extracted by maceration appeared as multiple protein bands, relative to the other venom sample. Low-molecular-weight proteins, revealed by western blotting, played an important immunogenic role in the production of antivenom. Lethality and protein levels varied according to the extraction method; venom obtained by the maceration technique showed lower toxicity than that obtained by electric stimulation.

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          Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4

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            Probit analysis

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              Blue silver: a very sensitive colloidal Coomassie G-250 staining for proteome analysis.

              A modified Neuhoff's colloidal Coomassie Blue G-250 stain is reported, dubbed "blue silver" on account of its considerably higher sensitivity, approaching the one of conventional silver staining. The main modifications, as compared to Neuhoff's protocol, were: a 20% increment in dye concentration (from 0.1% up to 0.12%) and a much higher level of phosphoric acid in the recipe (from 2% up to 10%). The "blue silver" exhibits a much faster dye uptake (80% during the first hour of coloration, vs. none with a commercial preparation from Sigma). Even at equilibrium (24 h staining), the "blue silver" exhibits a much higher sensitivity than all other recipes, approaching (but lower than) the one of the classical silver stain. Measurements of stain sensitivity after sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of bovine serum albumin (BSA) gave a detection limit (signal-to-noise ratio > 3) of 1 ng in a single zone. The somewhat lower sensitivity of "blue silver" as compared to classical silvering protocols in the presence of aldehydes is amply compensated for by its full compatibility with mass spectrometry of eluted polypeptide chains, after a two-dimensional map analysis, thus confirming that no dye is covalently bound (or permanently modifies) to any residue in the proteinaceous material. It is believed that the higher level of phosphoric acid in the recipe, thus its lower final pH, helps in protonating the last dissociated residues of Asp and Glu in the polypeptide coils, thus greatly favoring ionic anchoring of dye molecules to the protein moiety. Such a binding, though, must be followed by considerable hydrophobic association with the aromatic and hydrophobic residues along the polypeptide backbone.
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                Author and article information

                Journal
                jvatitd
                Journal of Venomous Animals and Toxins including Tropical Diseases
                J. Venom. Anim. Toxins incl. Trop. Dis
                Centro de Estudos de Venenos e Animais Peçonhentos (Botucatu, SP, Brazil )
                1678-9199
                2007
                : 13
                : 4
                : 844-856
                Affiliations
                [03] Ankara orgnameHacettepe University orgdiv1Faculty of Sciences orgdiv2Department of Statistics Turkey
                [01] Ankara orgnameRefik Saydam Hygiene Center Turkey
                [02] Ankara orgnameFaculty of Veterinary Medicine orgdiv1Department of Entomology Turkey
                Article
                S1678-91992007000400013 S1678-9199(07)01300413
                10.1590/S1678-91992007000400013
                13d92047-00df-4fc0-a7de-04257b49c8e4

                This work is licensed under a Creative Commons Attribution 4.0 International License.

                History
                : 06 March 2007
                : 09 May 2007
                Page count
                Figures: 0, Tables: 0, Equations: 0, References: 31, Pages: 13
                Product

                SciELO Brazil

                Self URI: Full text available only in PDF format (EN)
                Categories
                Original Papers

                gland maceration,immunogenicity,electric stimulation,lethality,scorpion venoms,Androctonus crassicauda

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