Characterization of 2':3'-cyclic nucleotide 3'-phosphodiesterase: rapid isolation, native enzyme analysis, identification of a serum-soluble activity, and kinetics.
The enzyme 2':3'-cyclic nucleotide 3'-phosphodiesterase (CNPase) was isolated from bovine brain white matter by a rapid (72 h) procedure. The minimum molecular weight (MW) of the enzyme was approximately 52,500 as estimated by sucrose density gradient analysis. When this isolated enzyme was stimulated with bovine serum albumin (BSA), the peak of activity was shifted to approximately 90,000 MW. Prior treatment by trypsin blocked the expression of the higher MW form of CNPase, but not the BSA activation of the enzyme. If the trypsin digestion was allowed to progress, the MW was gradually lowered to a broad peak sedimenting between 20,000 and 50,000 MW. An apparently soluble form of CNPase found in serum is described. Kinetic and MW comparisons between the serum soluble enzyme and CNPase isolated from bovine brain, as well as an analysis of substrate specificity, were made and it was concluded that the two enzymes were identical.
D015087:2',3'-Cyclic-Nucleotide Phosphodiesterases / Q000097:blood / Q000302:isolation & purification*; D000818:Animals; D001921:Brain / Q000201:enzymology*; D002417:Cattle; D002499:Centrifugation, Density Gradient; D006801:Humans; D006868:Hydrolysis; D007700:Kinetics; D008970:Molecular Weight; D010727:Phosphoric Diester Hydrolases / Q000302:isolation & purification*; D011817:Rabbits; D012710:Serum Albumin, Bovine / Q000494:pharmacology; D013379:Substrate Specificity; D014357:Trypsin / Q000378:metabolism
D012710:Serum Albumin, Bovine; D015087:2',3'-Cyclic-Nucleotide Phosphodiesterases; D010727:Phosphoric Diester Hydrolases; D014357:Trypsin