Isolation, characterization and Ca2+ and calmodulin regulation of cyclic nucleotide phosphodiesterases from human placentae of different ages.
Two kinds of phosphodiesterases were isolated from human placenta by DEAE chromatography and characterized: one Ca2+ and calmodulin dependent, the other stimulated by Ca2+ but not by calmodulin. Both hydrolyzed cAMP and cGMP. The first one exhibited a higher affinity for cGMP. Half maximal activation by calmodulin was attained at 10(-8)M of calmodulin concentration independently of the hydrolyzed substrate (cGMP or cAMP). This phosphodiesterase appears to be almost homogeneous by molecular sieve chromatography on Ultragel AcA 34. The second phosphodiesterase exhibited similar affinities for cAMP and cGMP and could be resolved into three active isoforms with different molecular weight on Ultrogel AcA 34. Only minor differences were observed in the characteristics of these enzymes when the phosphodiesterases were prepared from placentae of 7-8 weeks of pregnancy or from normal term placenta.
Tertrin-Clary
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C
Lab. des Hormones Polypeptidiques, CNRS, Gif Sur Yvette Cedex, INSERM Unité.
Chenut
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MC
Acker
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G
De La Llosa
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P
D015105:3',5'-Cyclic-AMP Phosphodiesterases / Q000302:isolation & purification / Q000378:metabolism*; D015106:3',5'-Cyclic-GMP Phosphodiesterases / Q000302:isolation & purification / Q000378:metabolism*; D002118:Calcium / Q000494:pharmacology*; D002147:Calmodulin / Q000494:pharmacology*; D002848:Chromatography, DEAE-Cellulose; D054677:Cyclic Nucleotide Phosphodiesterases, Type 1; D005260:Female; D006801:Humans; D007527:Isoenzymes / Q000302:isolation & purification / Q000378:metabolism*; D007700:Kinetics; D010920:Placenta / Q000201:enzymology* / Q000502:physiology; D011247:Pregnancy; D011261:Pregnancy Trimester, First; D011263:Pregnancy Trimester, Third; D013379:Substrate Specificity
D002147:Calmodulin; D007527:Isoenzymes; D015105:3',5'-Cyclic-AMP Phosphodiesterases; D054677:Cyclic Nucleotide Phosphodiesterases, Type 1; D015106:3',5'-Cyclic-GMP Phosphodiesterases; D002118:Calcium