SIMPLE DIRECT ASSAY OF 3',5'-CYCLIC NUCLEOTIDE PHOSPHODIESTERASE ACTIVITY BASED ON THE USE OF POLYACRYLAMIDE-BORONATE AFFINITY GEL CHROMATOGRAPHY
- 1 January 1979
- journal article
- research article
- Vol. 5 (1) , 65-74
Abstract
A rapid, simple, and direct assay for rat brain 3'',5''-cyclic nucleotide phosphodiesterase activity is based on the effective separation of cyclic(c)AMP, cGMP or cCMP from their corresponding 5''-nucleotides and nucleosides by chromatography on a polyacrylamide-boronate gel. The affinity of the boronate residue for cis-diols results in the retention of 5''-nucleotides and nucleosides while 3'',5''-cyclic nucleotides are not retained. The coelution of all 5''-nucleotides and nucleosides allows for the accurate assessment of phosphodiesterase activity in preparations contaminated by other purine metabolizing enzymes such as 5''-nucleotidases and nucleotide and nucleoside deaminases. Phosphodiesterase activity assayed by this means yields linear reaction kinetics with respect to time and amount of enzyme protein. Low blank values obtained allow for detection of as little as 2-3% conversion of substrate to product.This publication has 9 references indexed in Scilit:
- Purification and characterization of guanosine 3':5'-monophosphate-specific phosphodiesterase from guinea pig lung.Journal of Biological Chemistry, 1977
- Assay of adenylate cyclase by use of polyacrylamide-boronate gel columnsAnalytical Biochemistry, 1977
- CHARACTERIZATION OF A SPECTROPHOTOMETRIC ASSAY FOR CAMP PHOSPHODIESTERASE1977
- Isolation of catecholamines in biological fluids by boric acid gelAnalytical Biochemistry, 1977
- Assay errors of cyclic 3′,5′-nucleotide phosphodiesterase activity based on the recovery of adenosine alone using an anionic-exchange resin columnAnalytical Biochemistry, 1976
- Ontogenetic changes in levels of phosphodiesterase for adenosine 3′:5′-monophosphate and guanosine 3′:5′-monophosphate in the lung, liver, brain and heart from guinea pigsBiochimica et Biophysica Acta (BBA) - General Subjects, 1976
- Quantitative estimation of 3′5′ cyclic AMP phosphodiesterase using anion exchange resin in a batch processAnalytical Biochemistry, 1976
- ADENOSINE 3',5'-PHOSPHATE IN BIOLOGICAL MATERIALS .1. PURIFICATION AND PROPERTIES OF CYCLIC 3',5'-NUCLEOTIDE PHOSPHODIESTERASE AND USE SF THIS ENZYME TO CHARACTERIZE ADENOSINE 3',5'-PHOSPHATE IN HUMAN URINE1962
- PROTEIN MEASUREMENT WITH THE FOLIN PHENOL REAGENTJournal of Biological Chemistry, 1951