Cyclic nucleotide phosphodiesterase PDE1C1 in human cardiac myocytes.
Isoforms in the PDE1 family of cyclic nucleotide phosphodiesterases were recently found to comprise a significant portion of the cGMP-inhibited cAMP hydrolytic activity in human hearts. We examined the expression of PDE1 isoforms in human myocardium, characterized their catalytic activity, and quantified their contribution to cAMP hydrolytic and cGMP hydrolytic activity in subcellular fractions of this tissue. Western blotting with isoform-selective anti-PDE1 monoclonal antibodies showed PDE1C1 to be the principal isoform expressed in human myocardium. Immunohistochemical analysis showed that PDE1C1 is distributed along the Z-lines and M-lines of cardiac myocytes in a striated pattern that differs from that of the other major dual-specificity cyclic nucleotide phosphodiesterase in human myocardium, PDE3A. Most of the PDE1C1 activity was recovered in soluble fractions of human myocardium. It binds both cAMP and cGMP with K(m) values of approximately 1 microm and hydrolyzes both substrates with similar catalytic rates. PDE1C1 activity in subcellular fractions was quantified using a new PDE1-selective inhibitor, IC295. At substrate concentrations of 0.1 microm, PDE1C1 constitutes the great majority of cAMP hydrolytic and cGMP hydrolytic activity in soluble fractions and the majority of cGMP hydrolytic activity in microsomal fractions, whereas PDE3 constitutes the majority of cAMP hydrolytic activity in microsomal fractions. These results indicate that PDE1C1 is expressed at high levels in human cardiac myocytes with an intracellular distribution distinct from that of PDE3A and that it may have a role in the integration of cGMP-, cAMP- and Ca(2+)-mediated signaling in these cells.
Vandeput
Fabrice
F
Cardiology Section, Veterans Affairs Salt Lake City Health Care System, 500 Foothill Boulevard, Salt Lake City, UT 84148, USA.
Wolda
Sharon L
SL
Krall
Judith
J
Hambleton
Ryan
R
Uher
Lothar
L
McCaw
Kim N
KN
Radwanski
Przemyslaw B
PB
Florio
Vincent
V
Movsesian
Matthew A
MA
D000818:Animals; D002460:Cell Line; D000242:Cyclic AMP / Q000378:metabolism; D006152:Cyclic GMP / Q000378:metabolism; D054677:Cyclic Nucleotide Phosphodiesterases, Type 1 / Q000235:genetics / Q000378:metabolism*; D015971:Gene Expression Regulation, Enzymologic; D006801:Humans; D006868:Hydrolysis; D007527:Isoenzymes / Q000378:metabolism; D007700:Kinetics; D032383:Myocytes, Cardiac / Q000201:enzymology*; D018411:Spodoptera; D013347:Subcellular Fractions / Q000201:enzymology
D007527:Isoenzymes; D000242:Cyclic AMP; D054677:Cyclic Nucleotide Phosphodiesterases, Type 1; C517272:PDE1C protein, human; D006152:Cyclic GMP