DOI: 10.1111/j.1471-4159.1979.tb05257.x. PURIFICATION OF 2′,3′‐CYCLIC NUCLEOTIDE 3′‐PHOSPHOHYDROLASE FROM BOVINE BRAIN BY IMMUNOAFFINITY CHROMATOGRAPHY: FURTHER BIOCHEMICAL CHARACTERIZATION OF THE PROTEIN

PURIFICATION OF 2′,3′‐CYCLIC NUCLEOTIDE 3′‐PHOSPHOHYDROLASE FROM BOVINE BRAIN BY IMMUNOAFFINITY CHROMATOGRAPHY: FURTHER BIOCHEMICAL CHARACTERIZATION OF THE PROTEIN

10.1111/j.1471-4159.1979.tb05257.x

Crossref journal-article

Wiley

Journal of Neurochemistry (311)

Abstract

Abstract— The purification of small amounts of 2′,3′‐cyclic nucleotide 3′‐phosphohydrolase from bovine white matter by ion‐exchange techniques (Drummondet al., 1978) has been used to provide antigen for the production of specific rabbit antibodies to this enzyme. Specific antibody has been purified from immune serum by affinity chromatography on a column of Sepharose to which the enzyme has been attached, and the purified antibody has been coupled to cyanogen bromide‐activated Sepharose. Affinity chromatography on the immunoadsorbent effectively purifies 2′,3′‐cyclic nucleotide 3 ‐phosphohydrolase in one step from an extract of an acetone powder made from bovine white matter. This modified purification procedure has reduced the time required for purification and increased the yield of the enzyme to 57%. In SDS‐gel electrophoresis in phosphate buffer the enzyme migrates as an aggregate of about 98,000MW. When the buffer is Tris‐glycine, the apparent MW is about 44,000 and under specific conditions two proteins of only slightly different mobilities can be discerned. Within experimental error the amino acid compositions of the proteins in the two bands are indistinguishable. Peptide patterns obtained by polyacrylamide gel electrophoresis following proteolytic digestion with Straphylococcus aureus V8 protease or papain show extensive structural homology between the two proteins, but detectable differences are apparent.

Bibliography

Drummond, R. J. (1979). PURIFICATION OF 2â²,3â²âCYCLIC NUCLEOTIDE 3â²âPHOSPHOHYDROLASE FROM BOVINE BRAIN BY IMMUNOAFFINITY CHROMATOGRAPHY: FURTHER BIOCHEMICAL CHARACTERIZATION OF THE PROTEIN. Journal of Neurochemistry, 33(6), 1143â1150. Portico.

Authors 2

Robert J. Drummond (first)
(additional)

References 26 Referenced 24

10.1016/0019-2791(69)90179-7
10.1016/0006-8993(72)90144-8
10.1016/S0021-9258(19)75212-0
10.1016/0006-291X(78)90907-5
10.1016/S0021-9258(19)70852-7 / J. biol. Chem. / Hydrolysis of ribonucleoside 2′,3′‐cyclic phosphates by a diesterase from brain by Drummond G. I. (1962)
10.1111/j.1471-4159.1978.tb00122.x
{'key': 'e_1_2_2_8_1', 'first-page': '410', 'article-title': 'Cytochemical localization of 2′,3′‐cyclic nucleotide 3′‐phosphohydrolase in CNS white matter', 'volume': '2', 'author': 'Encel E. L.', 'year': '1976', 'journal-title': 'Neurosci. Abstr.'} / Neurosci. Abstr. / Cytochemical localization of 2′,3′‐cyclic nucleotide 3′‐phosphohydrolase in CNS white matter by Encel E. L. (1976)
10.1073/pnas.67.3.1425
10.1073/pnas.74.3.864
10.1016/0003-2697(71)90348-4
10.1016/0006-8993(73)90832-9
10.1016/0009-8981(66)90114-8
10.1111/j.1471-4159.1967.tb06164.x
10.1111/j.1471-4159.1968.tb10329.x
10.1111/j.1471-4159.1970.tb02252.x
10.1038/227680a0
10.1016/S0021-9258(19)52451-6
10.1139/o69-151
10.1111/j.1699-0463.1953.tb00247.x
10.1073/pnas.68.4.770
10.1111/j.1471-4159.1975.tb03842.x
10.1111/j.1471-4159.1972.tb01388.x
10.1111/j.1471-4159.1978.tb10779.x
10.1073/pnas.72.5.1927
10.1016/S0021-9258(18)94333-4 / J. biol. Chem. / Reliability of molecular weight determinations by dodecyl sulfate polyacrylamide gel electrophoresis by Weber K. (1969)
10.1111/j.1471-4159.1972.tb01401.x

Dates

Type	When
Created	18 years, 10 months ago (Oct. 5, 2006, 6:06 a.m.)
Deposited	1 year, 9 months ago (Nov. 4, 2023, 9:01 a.m.)
Indexed	1 year ago (Aug. 6, 2024, 3:59 a.m.)
Issued	45 years, 8 months ago (Dec. 1, 1979)
Published	45 years, 8 months ago (Dec. 1, 1979)
Published Online	18 years, 10 months ago (Oct. 5, 2006)
Published Print	45 years, 8 months ago (Dec. 1, 1979)

Funders 0

None

BibTeX

@article{Drummond_1979, title={PURIFICATION OF 2′,3′‐CYCLIC NUCLEOTIDE 3′‐PHOSPHOHYDROLASE FROM BOVINE BRAIN BY IMMUNOAFFINITY CHROMATOGRAPHY: FURTHER BIOCHEMICAL CHARACTERIZATION OF THE PROTEIN}, volume={33}, ISSN={1471-4159}, url={http://dx.doi.org/10.1111/j.1471-4159.1979.tb05257.x}, DOI={10.1111/j.1471-4159.1979.tb05257.x}, number={6}, journal={Journal of Neurochemistry}, publisher={Wiley}, author={Drummond, Robert J.}, year={1979}, month=dec, pages={1143–1150} }

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