Mingone, Christopher J., Sachin A. Gupte, Joseph L. Chow, Mansoor Ahmad, Nader G. Abraham, and Michael S. Wolin. Protoporphyrin IX generation from ␦-aminolevulinic acid elicits pulmonary artery relaxation and soluble guanylate cyclase activation. Am J Physiol Lung Cell Mol Physiol 291: L337-L344, 2006; doi:10.1152/ajplung.00482.2005.-Protoporphyrin IX is an activator of soluble guanylate cyclase (sGC), but its role as an endogenous regulator of vascular function through cGMP has not been previously reported. In this study we examined whether the heme precursor ␦-aminolevulinic acid (ALA) could regulate vascular force through promoting protoporphyrin IX-elicited activation of sGC. Exposure of endothelium-denuded bovine pulmonary arteries (BPA) in organoid culture to increasing concentrations of the heme precursor ALA caused a concentration-dependent increase in BPA epifluorescence, consistent with increased tissue protoporphyrin IX levels, associated with decreased force generation to increasing concentrations of serotonin. The force-depressing actions of 0.1 mM ALA were associated with increased cGMP-associated vasodilator-stimulated phosphoprotein (VASP) phosphorylation and increased sGC activity in homogenates of BPA cultured with ALA. Increasing iron availability with 0.1 mM FeSO 4 inhibited the decrease in contraction to serotonin and increase in sGC activity caused by ALA, associated with decreased protoporphyrin IX and increased heme. Chelating endogenous iron with 0.1 mM deferoxamine increased the detection of protoporphyrin IX and force depressing activity of 10 M ALA. The inhibition of sGC activation with the heme oxidant 10 M 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ) attenuated the force depressing actions of an NO donor without altering the actions of ALA. Thus control of endogenous formation of protoporphyrin IX from ALA by the availability of iron is potentially a novel physiological mechanism of controlling vascular function through regulating the activity of sGC.guanosine 3Ј,5Ј-cyclic monophosphate; heme metabolism; iron; vasodilation THE REGULATION OF THE SOLUBLE FORM of guanylate cyclase (sGC) by nitric oxide (NO) has been extensively studied for its role in promoting relaxation in the pulmonary and systemic vasculature through increasing production of cGMP (2,12,23,33). The bovine pulmonary arteries (BPA) examined in this study show an endothelium-dependent relaxation to NO, which is mediated through the stimulation of sGC (17,18). NO relaxes BPA through stimulating sGC in a manner that is controlled by a thiol redox mechanism regulated by cytosolic NADPH and glutathione redox (25). Under hypoxia the mechanism of relaxation to NO appears to shift to a cGMPindependent mechanism stimulating calcium-reuptake by the sarco(endo)plasmic reticulum Ca 2ϩ -adenosinetriphosphatase (SERCA) pump (26). NO activates sGC by binding a ferrous heme group, which appears to be a cofactor that is normally bound to sGC when it is isolated from tissues (6, 9, 34). Early studies investigating how NO activate...