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Identification of Cytosolic Phosphodiesterases in the Erythrocyte: A Possible Role for PDE5

Identification of Cytosolic Phosphodiesterases in the Erythrocyte: A Possible Role for PDE5

Shaquria P Adderley, Kelly M Thuet, Meera Sridharan, Elizabeth A Bowles, Alan H Stephenson, Mary L Ellsworth, Randy S Sprague

Med Sci Monit. 2011 May;17(5):CR241-7.

PMID: 21525805

Abstract:

Background:
Within erythrocytes (RBCs), cAMP levels are regulated by phosphodiesterases (PDEs). Increases in cAMP and ATP release associated with activation of β-adrenergic receptors (βARs) and prostacyclin receptors (IPRs) are regulated by PDEs 2, 4 and PDE 3, respectively. Here we establish the presence of cytosolic PDEs in RBCs and determine a role for PDE5 in regulating levels of cGMP.
Material/methods:
Purified cytosolic proteins were obtained from isolated human RBCs and western analysis was performed using antibodies against PDEs 3A, 4 and 5. Rabbit RBCs were incubated with dbcGMP, a cGMP analog, to determine the effect of cGMP on cAMP levels. To determine if cGMP affects receptor-mediated increases in cAMP, rabbit RBCs were incubated with dbcGMP prior to addition of isoproterenol (ISO), a βAR receptor agonist. To demonstrate that endogenous cGMP produces the same effect, rabbit and human RBCs were incubated with SpNONOate (SpNO), a nitric oxide donor, and YC1, a direct activator of soluble guanylyl cyclase (sGC), in the absence and presence of a selective PDE5 inhibitor, zaprinast (ZAP).
Results:
Western analysis identified PDEs 3A, 4D and 5A. dbcGMP produced a concentration dependent increase in cAMP and ISO-induced increases in cAMP were potentiated by dbcGMP. In addition, incubation with YC1 and SpNO in the presence of ZAP potentiated βAR-induced increases in cAMP.
Conclusions:
PDEs 2, 3A and 5 are present in the cytosol of human RBCs. PDE5 activity in RBCs regulates cGMP levels. Increases in intracellular cGMP augment cAMP levels. These studies suggest a novel role for PDE5 in erythrocytes.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
IAR42416687	Anti-phospho-FOXO1A/3A (pSer322+Ser325) antibody produced in rabbit			Price
IAR4248237	Anti-α2C Adrenergic Receptor antibody produced in rabbit			Price

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