Analysis of nitric oxide-sensitive guanylyl cyclase in human platelets before and after aggregation

Nitric oxide (NO) inhibits platelet adhesion to vascular endothelium and platelet aggregation through activation of soluble guanylyl cyclase (sGC) and a consequent increase in cGMP. The aim of the present study was to analyze NO-sensitive sGC in human platelets before and after aggregation. NO-sensitive sGC activity was tested in the cytosol and membrane fractions of native human platelets and ADP-induced platelet aggregates in the presence of 3 mM Mn2+ as cofactor. After ADP-induced platelet aggregation there was a significant increase of sGC activity in membranes. Western blot analysis showed a partial translocation of the enzyme to the plasma membrane. These findings support recent data that sGC is associated with cellular membranes in various tissues and cell types and that this membrane association is influenced by the activation state in human platelets (Nat Cell Biol 2002; 4: 307–11). Using 3 mM Mg2+ instead of Mn2+ as cofactor, a sharp decrease of sGC activity was apparent in the cytosol of aggreg...