Abstract
A simple method for the amay of adenyl cyclase in tissue homogenates is described that uses either α-P82-, C14-or H3-labeled adenosine triphosphate (ATP) as substrate. Cyclic 3’, 5’- adenosine monophosphate (cyclic 3’, S’-AMP) formed during the incubation was isolated by chromatography on Dowex 50-H+ columns followed by precipitation of all nucleotides and inorganic phosphates by ZnSO4-Ba(OH)2, treatment, which left cyclic 3’, S’-AMP in the supernatant fluid. The purity of the cyclic 3’, 5’-AMP fraction was determined with the use of ion-exchange chromatography and paper chromatographic and electrophoretic techniques, as well as crystallization to constant specific activity. Furthermore, the purity of the nucleotide has been assessed enzymatically, by utilizing a purified cyclic nucleotide phosphodiesterase. The method described here makes possible the measurement of enzyme activities in tissue weighing less than 1 mg. Moreover, the entire procedure can be completed less than 3 hr.
Footnotes
- Received March 25, 1968.
- Accepted May 28, 1968.
- © 1968 by the Williams & Wilkins Co.
JPET articles become freely available 12 months after publication, and remain freely available for 5 years.Non-open access articles that fall outside this five year window are available only to institutional subscribers and current ASPET members, or through the article purchase feature at the bottom of the page.
|
Log in using your username and password
Purchase access
In this issue
A SIMPLE, SENSITIVE METHOD FOR THE ASSAY OF ADENYL CYCLASE
