A purification of venom phosphodiesterase.
نویسندگان
چکیده
For purposes of analysis of polynucleotides, it is desirable to have a phosphodiesterase, substantially free of 5-nucleotidase or other phosphatase activity. The presence of a phosphodiesterase in a wide variety of snake venoms was demonstrated by Gullan<l and Jackson (1). These venoms were found also to contain a potent 5-nucleotidase, but were free of alkaline phosphatase activity. Hurst and Butler (2) found that certain samples of Russell's viper venom were nearly free of 5-nucleotidase activity, while retaining potent phosphodiesterase action. By a chromatographic procedure, involving the use of cellulose columns, they were able to reduce the 5-nucleotidase activity of rattlesnake venom, relative to its phosphodiesterase activity, and to obtain fractions nearly comparable to the viper venom. For certain purposes, however, such as the determination of the ratio of sequential isomers in mixed isomeric dinucleotides, the preparations obtained from rattlesnake venom by the chromatographic procedure of Hurst and Butler are not sufficiently free of 5-nucleotidase action. Therefore, a simple acetone fractionation has been developed, which yields a potent phosphodiesterase, substantially free of 5-nucleotidase activity, from rattlesnake venom. With this preparation, it has been possible to obtain quantitative degradation of diand trinucleotides to mononucleotides, and to degrade a complete desoxyribonuclease digest to mononucleotides with the conversion of only 1 per cent of the digest to nucleosides.
منابع مشابه
Purification of phosphodiesterase from Bothrops atrox venom, with special consideration of the elimination of monophosphatases.
In fragments of nucleic acids bearing 3’-monophosphoryl groups, both terminals may be determined by degrading the fragment with a massive dose of a single enzyme, venom phosphodiesterase (l-5). To be reliable, this method requires a preparation of phosphodiesterase which is both highly potent and free from contaminating interfering enzymes. The most dangerous contaminant is venom endonuclease, ...
متن کاملA Specific and Nonspecific Alkaline Monophosphatase in the Venom of Bothrops atrox and Their Occurrence in the Purified Venom
Reis (1) discovered 5’-nucleotidase in 1934. Shortly thereafter, Gulland and Jackson (2) showed its presence in venoms of many species of snake. Although several methods of separation of 5’-nucleotidase from phosphodiesterase of venom have been proposed (3-7), the available preparations of 5’-nucleotidase are still rather crude (8-10). Two reasons prompted us to purify venom 5’-nucleotidase. Fi...
متن کاملA specific and nonspecific alkaline monophosphatase in the venom of Bothrops atrox and their occurrence in the purified venom phosphodiesterase.
Reis (1) discovered 5’-nucleotidase in 1934. Shortly thereafter, Gulland and Jackson (2) showed its presence in venoms of many species of snake. Although several methods of separation of 5’-nucleotidase from phosphodiesterase of venom have been proposed (3-7), the available preparations of 5’-nucleotidase are still rather crude (8-10). Two reasons prompted us to purify venom 5’-nucleotidase. Fi...
متن کاملPartial Purification and Characterization of Anticoagulant Factor from the Snake (Echis carinatus) Venom
Objective(s): Snake venoms contain complex mixture of proteins with biological activities. Some of these proteins affect blood coagulation and platelet function in different ways. Snake venom toxin may serve as a starting material for drug design to combat several pathophysiological problems such as cardiovascular disorders. In the present study, purification of anticoagulation facto...
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عنوان ژورنال:
- The Journal of biological chemistry
دوره 198 1 شماره
صفحات -
تاریخ انتشار 1952