Isolation of 2',3'-Cyclic Nucleotide 3'-Phosphodiesterase from Human Brain |
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Authors: | Terry Joe Sprinkle Marilyn J Grimes A Gary Eller |
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Institution: | Vetertins Administration Medical Center and Medical College of Georgia, Augustu, Georgia 30904, U.S.A. |
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Abstract: | Abstract: 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. |
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