Purification of phosphodiesterase from Bothrops atrox venom, with special consideration of the elimination of monophosphatases.
نویسنده
چکیده
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, which was recently purified (3). Fortunately, the optimal pH of the endonuclease is 5, and the enzyme has almost no activity at pH 8.9, the optimum for phosphodiesterase. Equally fortunate are the differences in charges between the two enzymes that permit their rather effective separation on either carboxymethylor diethylaminoethyl cellulose. The preceding paper (5) describes the purification and properties of the two monophosphatases which contaminate venom phosphodiesterase. In the earlier methods of preparation of phosphodiesterase (4, 6-8), the major effort was directed toward the removal of 5’-nucleotidase, but the nonspecific phosphatase was totally neglected. This paper describes a modified procedure for the preparation of phosphodiesterase. The procedure was aimed at obtaining phosphodiesterase free from both contaminating monophosphatases. This objective was achieved in respect to 5’-nucleotidase but only partly attained in respect to the nonspecific phosphatase.
منابع مشابه
A 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...
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عنوان ژورنال:
- The Journal of biological chemistry
دوره 238 شماره
صفحات -
تاریخ انتشار 1963