Endothelial-derived nitric oxide (NO) is an important intercellular messenger. Although endothelial cells contain both nitric oxide synthase and soluble guanylate cyclase, the nature of receptor proteins for cGMP is uncertain. Based on previous work in vascular smooth muscle cells which indicates that the cGMP-dependent protein kinase (cGK) is partially associated with the cytoskeleton, we determined that cGK was present in non-cytosolic fractions of endothelial cells. The data reveal that cGK is found only in Triton-soluble extracts of particulate fractions from bovine aortic endothelial cells and provide the first evidence for the existence of cGK in this cell type based on immunoreactivity, immunofluorescence microscopy and phosphotransferase activity. The limited distribution of endothelial cell cGK may explain why this kinase has not been heretofore identified in endothelial cells. © 1994 Academic Press, Inc.