Radioactive measurement of 2' ,3'-cyclic nucleotide 3'-phosphodiesterase activity in the central and peripheral nervous system and in extraneural tissue

Sprinkle, T.J.; Zaruba, M.E.; McKhann, G.M.

Analytical Biochemistry 88(2): 449-456


ISSN/ISBN: 0003-2697
PMID: 211873
DOI: 10.1016/0003-2697(78)90443-8
Accession: 068524246

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A new, sensitive and reproducible method for the determination of 2',3'-cyclic-nucleotide 3'-phosphodiesterase (EC activity in both central and peripheral nervous system tissue and in extra-neural tissue is described. [8-3H]2',3'-cyclic AMP was used as the substrate. The [8-3H]2'-AMP product formed was isolated by TLC or, alternatively, the reaction was coupled with an excess of Escherichia coli alkaline phosphatase, and the [8-3H]adenosine formed was isolated by column chromatography on AG 1-X2 resin. The values obtained by the 2 methods were compared with those obtained using a spectrophotometric method. Hydrolysis rates of 0.50 nmol/min were reproducibly measured in 18-day fetal rat spinal cord.