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Characterization of cyclic nucleotide phosphodiesterase activity in Phaseolus vulgaris
Author(s) -
Dupon M.,
Onckelen H. A.,
Greef J. A.
Publication year - 1987
Publication title -
physiologia plantarum
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.351
H-Index - 146
eISSN - 1399-3054
pISSN - 0031-9317
DOI - 10.1111/j.1399-3054.1987.tb04301.x
Subject(s) - chromatofocusing , phaseolus , chromatography , chemistry , gel permeation chromatography , cyclic nucleotide phosphodiesterase , agarose , high performance liquid chromatography , enzyme assay , phosphodiesterase , enzyme , ion chromatography , biochemistry , isoelectric point , biology , organic chemistry , botany , polymer
Cyclic nucleotide phosphodiesterase (3′,5′‐cyclic nucleotide nucleotidohydrolase, EC 3.1.4.17) activity isolated from Phaseolus vulgaris L. cv. Limberg seedlings was partially purified and characterized by fractional (NH 4 ) 2 SO 4 precipitation, DEAE‐cellulose chromatography, chromatography on 3′,5′‐cAMP‐agarose, gel permeation chromatography and chromatofocusing. A crude enzyme preparation, a 30–65% (NH 4 ) 2 SO 4 pellet, showed an acidic pH optimum. The enzyme activity was stimulated by imidazole and divalent cations such as Ca 2+ , Mg 2+ and Mn 2+ , whereas NaF, PP i and Fe 3+ were inhibitory. Isobutylmethylxanthine had no significant effect on the plant enzyme. An M I of 42 000 was estimated by gel permeation high performance liquid chromatography. By chromatography on 3′,5′‐cAMP‐agarose a phosphodiesterase was resolved that produced 5′‐AMP as sole reaction product.