Purification and characterization of high-affinity cyclic adenosine 5'-monophosphate phosphodiesterases from human acute myelogenous leukemic cells

P. Onali, S. J. Strada, L. Chang, P. M. Epstein, Evan M Hersh, W. J. Thompson

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

Soluble, high-affinity cyclic adenosine 3':5'-monophosphate (cyclic AMP) phosphodiesterases extracted from blast cells of patients with acute myelogenous leukemia have been characterized by physical, kinetic, and immunological criteria and fractionated to a high degree of purity. Procedures used in this study were similar to those used to purify the high-affinity enzyme from dog kidney. Two forms of high-affinity enzyme were found in blast cells. Form A was similar to the known type IV phosphodiesterases, including those of normal lymphocytes and monocytes. It showed a molecular weight near 60,000, a rate of hydrolysis of cyclic AMP 7 to 10 times that of cyclic guanosine 3':5'-monophosphate (cyclic GMP), competitive inhibition by cyclic GMP for cyclic AMP hydrolysis, and identical immunoreactivity by Western transfer analysis. This enzyme form was purified to apparent homogeneity by physical criteria but showed a low maximum velocity relative to other phosphodiesterase forms. A second, different form of high-affinity phosphodiesterase (Form B) was also resolved and partially purified. By comparison with Form A, this enzyme eluted from diethylaminoethyl cellulose at slightly lower ionic strength, had a lower molecular weight, appeared specific for cyclic AMP as substrate, showed no inhibition of cyclic AMP hydrolysis by cyclic GMP, and displayed no immunological cross-reactivity to the M(r) 60,000 enzyme. Neither enzyme form was activated by calmodulin or proteolysis, whereas both showed comparable inhibition by 6,7-dimethoxy-1-veratrylisoquinoline, 1-methyl-3-isobutylxanthine, and 1,3-dimethylxanthine.

Original languageEnglish (US)
Pages (from-to)1384-1391
Number of pages8
JournalCancer Research
Volume45
Issue number3
StatePublished - 1985
Externally publishedYes

Fingerprint

Type 5 Cyclic Nucleotide Phosphodiesterases
Cyclic AMP
Guanosine Monophosphate
Phosphoric Diester Hydrolases
Enzymes
Hydrolysis
Molecular Weight
Type 4 Cyclic Nucleotide Phosphodiesterase
1-Methyl-3-isobutylxanthine
Calmodulin
Theophylline
Acute Myeloid Leukemia
Cellulose
Adenosine
Osmolar Concentration
Proteolysis
Monocytes
Dogs
Lymphocytes
Kidney

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

Purification and characterization of high-affinity cyclic adenosine 5'-monophosphate phosphodiesterases from human acute myelogenous leukemic cells. / Onali, P.; Strada, S. J.; Chang, L.; Epstein, P. M.; Hersh, Evan M; Thompson, W. J.

In: Cancer Research, Vol. 45, No. 3, 1985, p. 1384-1391.

Research output: Contribution to journalArticle

Onali, P. ; Strada, S. J. ; Chang, L. ; Epstein, P. M. ; Hersh, Evan M ; Thompson, W. J. / Purification and characterization of high-affinity cyclic adenosine 5'-monophosphate phosphodiesterases from human acute myelogenous leukemic cells. In: Cancer Research. 1985 ; Vol. 45, No. 3. pp. 1384-1391.
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AU - Epstein, P. M.

AU - Hersh, Evan M

AU - Thompson, W. J.

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