Hyaluronidase and esterase activities of the venom of the poisonous brown recluse spider
| Autor: | K.D. Elgert, James T. Barrett, R.P. Wright, Benedict J. Campbell |
|---|---|
| Rok vydání: | 1973 |
| Předmět: |
Immunodiffusion
Iron Guinea Pigs Antivenom Biophysics Hyaluronoglucosaminidase Venom complex mixtures Biochemistry Esterase chemistry.chemical_compound Hyaluronidase medicine Animals Carbon Radioisotopes Sodium dodecyl sulfate Molecular Biology Polyacrylamide gel electrophoresis Chelating Agents Gel electrophoresis Chromatography Antivenins Venoms Esterases Spiders Hydrogen-Ion Concentration Molecular Weight chemistry Sephadex Chromatography Gel Microscopy Electron Scanning Electrophoresis Polyacrylamide Gel Rabbits Chloromercuribenzoates Copper Phenanthrolines medicine.drug |
| Zdroj: | Archives of Biochemistry and Biophysics. 159:415-426 |
| ISSN: | 0003-9861 |
| Popis: | Venom of Loxosceles reclusa free from impurities was expressed from venom glands collected by microdissection. Polyacrylamide gel electrophoresis of the venom at pH 8.3 demonstrated 7 or 8 major plus 3 or 4 minor components. Upon electrophoresis at pH 4.9 two major components plus 3 or 4 minor components were noted. Monophoretic hyaluronidase prepared by Sephadex gel filtration and electrophoresis at pH 8.3 exhibited optimum activity from pH 5.0 to 6.6. Sodium dodecyl sulfate gel electrophoresis of purified hyaluronidase revealed two components with estimated molecular weights of 33,000 and 63,000. The purified hyaluronidase exhibited activity against chondroitin sulfate, types A, B, and C at approximately 20–30% of that upon hyaluronic acid. The enzyme was inhibited 10–20% by the heavy metal ions, Fe+3 and Cu+2. Rabbit antivenom inhibited the spreading effect of whole venom in vivo and completely inhibited hyaluronidase in vitro. Incorporation of [14C]leucine into the spider venom led to the separation of hyaluronidase from the dermonecrotic activity of the venom. The venom demonstrated activity against carbobenzoxy- l -tyrosine-p-nitrophenyl ester and β-naphthylacetate which was inhibited approximately 65% by 2.5 × 10−3 m levels of EDTA and EGTA but not by 2.5 × 10−4 m o-phenanthroline. The esterase activity resisted concentrations of p-chloromercuribenzoate which totally inactivated papain. The venom appeared devoid of collagenase, dipeptidase, acetylcholinesterase, phosphodiesterase, ribonuclease A, and deoxyribonuclease. |
| Databáze: | OpenAIRE |
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