Document Detail

Characterization of cyclic nucleotide phosphodiesterase isoforms associated to isolated cardiac nuclei.
MedLine Citation:
PMID:  10564757     Owner:  NLM     Status:  MEDLINE    
The identity and location of nuclear cyclic nucleotide phosphodiesterases (PDE) has yet to be ascertained. Intact cardiac nuclei and subnuclear fractions from ovine hearts were isolated to determine cAMP-specific PDE activity which was 3-fold greater than that of cGMP PDE, the latter being insensitive to Ca-calmodulin and zaprinast. Specific hydrolytic activities of the cardiac nuclear envelopes (NE) were similar to those measured in the corresponding intact nuclei, thus suggesting that most PDE activity is associated with the nuclear membrane. Moreover, the main hydrolytic activities in cardiac nuclei were attributed to PDE4 (56%) and PDE3 (44%). The pharmacological sensitivity of each isoform in terms of IC(50), K(m) and K(i) values was typical of previously characterized cardiac PDE 3 and 4 isoforms. PDE2 (cGMP-stimulated PDE) represented a minor component (8-9%) of total hydrolytic activity. Solubilization of nuclear envelopes and HPLC separation also yielded rolipram-sensitive PDE activities. Upon 1% Triton X-100 extractions, the presence of PDE3 and PDE4 was revealed in a low speed, nucleopore complex-enriched, P1 pellet. In addition, Western blot analysis demonstrated the presence of PDE4B and PDE4D subtypes in the nuclei as well as enrichment in NE. However, in the same preparations, the presence of PDE4A could not be ascertained. Altogether, these results suggest an intrinsic and predominant association of these nuclear PDEs with the NE and much likely with nucleopore complexes.
C Lugnier; T Keravis; A Le Bec; O Pauvert; S Proteau; E Rousseau
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Biochimica et biophysica acta     Volume:  1472     ISSN:  0006-3002     ISO Abbreviation:  Biochim. Biophys. Acta     Publication Date:  1999 Nov 
Date Detail:
Created Date:  2000-01-06     Completed Date:  2000-01-06     Revised Date:  2007-11-15    
Medline Journal Info:
Nlm Unique ID:  0217513     Medline TA:  Biochim Biophys Acta     Country:  NETHERLANDS    
Other Details:
Languages:  eng     Pagination:  431-46     Citation Subset:  IM    
Laboratoire de Pharmacologie et de Physico-chimie des Interactions Cellulaires et Moléculaires, CNRS-UMR, Université Louis Pasteur de Strasbourg, 67401, Illkirch, France.
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MeSH Terms
1-Methyl-3-isobutylxanthine / pharmacology
3',5'-Cyclic-AMP Phosphodiesterases / antagonists & inhibitors,  isolation & purification,  metabolism*
Cell Nucleus / enzymology
Chromatography, High Pressure Liquid
Cyclic Nucleotide Phosphodiesterases, Type 3
Cyclic Nucleotide Phosphodiesterases, Type 4
Heart Ventricles
Intracellular Membranes / enzymology
Isoenzymes / metabolism
Myocardium / enzymology*
Pyrazines / pharmacology
Rolipram / pharmacology
Reg. No./Substance:
0/Isoenzymes; 0/Pyrazines; 28822-58-4/1-Methyl-3-isobutylxanthine; 61413-54-5/Rolipram; 89541-55-9/5-(4-acetamidophenyl)pyrazin-2(1H)-one; EC',5'-Cyclic-AMP Phosphodiesterases; EC Nucleotide Phosphodiesterases, Type 3; EC Nucleotide Phosphodiesterases, Type 4

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