Gene/Protein
Disease
Symptom
Drug
Enzyme
Compound
Pivot Concepts:
Gene/Protein
Disease
Symptom
Drug
Enzyme
Compound
Target Concepts:
Gene/Protein
Disease
Symptom
Drug
Enzyme
Compound
Query: EC:3.1.4.37 (
CNPase
)
539
document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)
1. A spectrophotometric assay of
2':3'-cyclic nucleotide 3'-phosphodiesterase
(
EC 3.1.4.37
) based on the use of an acid-base indicator and a buffer having identical pKa values is described. The assay is simple and rapid; it was particularly convenient for monitoring the enzyme activity at various stages of purification. 2. Several proteinases were examined for their ability to solubilize
2':3'-cyclic nucleotide 3'-phosphodiesterase
from delipidated brain white matter.
Trypsin
(EC 3.4.21.4) and elastase (EC 3.4.21.11) appeared to be more effective than the other proteinases examined.
Trypsin
, however, caused inactivation; elastase was therefore chosen to solubilize
2':3'-cyclic nucleotide 3'-phosphodiesterase
. When a partially purified preparation of
2':3'-cyclic nucleotide 3'-phosphodiesterase
was treated with elastase,
2':3'-cyclic nucleotide 3'-phosphodiesterase
was solubilized nearly quantitatively. Elastatinal, a specific inhibitor of elastase, specifically inhibited the solubilization with elastase. 3.
2':3'-cyclic nucleotide 3'-phosphodiesterase
was purified from bovine brain white matter by: (i) delipidation; (ii) solubilization with hexadecyltrimethylammonium bromide; (iii) gel chromatography on Sepharose; (iv) ethanol precipitation and resolubilization by digestion with elastase; (v) chromatography on DEAE-Sephadex; (vi) affinity chromatography on 8-(6-aminohexyl)amino-2'-AMP-Sepharose. 4. The purified enzyme migrated as a single protein band on polyacrylamide-gel electrophoresis at pH 4.3 and on sodium dodecyl sulphate/polyacrylamide-gel electrophoresis; the estimated mol.wt. in the latter electrophoresis was 27000-31000. Gel filtration of the purified enzyme through Sephadex G-150 indicated a mol.wt. of 31000. Therefore the purified enzyme is a monomer protein with a mol.wt. of approx. 30000.
...
PMID:Spectrophotometric assay, solubilization and purification of brain 2':3'-cyclic nucleotide 3'-phosphodiesterase. 625 86
