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Isolation of 2′,3′‐Cyclic Nucleotide 3′‐Phosphodiesterase from Human Brain
Author(s) -
Sprinkle Terry Joe,
Grimes Marilyn J.,
Eller A. Gary
Publication year - 1980
Publication title -
journal of neurochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.75
H-Index - 229
eISSN - 1471-4159
pISSN - 0022-3042
DOI - 10.1111/j.1471-4159.1980.tb09661.x
Subject(s) - dithiothreitol , enzyme , phosphodiesterase , cyclic nucleotide phosphodiesterase , polyacrylamide gel electrophoresis , biochemistry , gel electrophoresis , acetone , nucleotide , chemistry , microbiology and biotechnology , specific activity , cyclic nucleotide , substrate (aquarium) , yield (engineering) , biology , ecology , materials science , metallurgy , gene
The enzyme 2′,3′‐cyclic nucleotide 3′‐phosphodiesterase (EC 3.1.4.37) has been isolated from an acetone powder of human subcortical white matter. The yield was about 11 mg from 28 g of powder and a specific activity of 213 unitdmg protein was obtained using 2′,3′‐cyclic CMP as the substrate. A major protein band of molecular weight approx. 96,000 was found by gel electrophoresis under nonreducing conditions. However, two distinct protein bands of molecular weight 46,000 ± 1400 and 48,000 ± 1400 were observed when the protein sample was reduced with 10 mM‐dithiothreitol and subjected to electrophoresis in more restrictive 12‐15% polyacrylamide‐SDS gels. This molecular weight is lower than that previously reported for the bovine enzyme. Antibodies against the purified human enzyme have been raised in New Zealand white rabbits.