The potent vasodilating and guanylyl cyclase activating dinitrosyl-iron(II) complex is stored in a protein-bound form in vascular tissue and is released by thiols.
We studied the biological activity, stability and interaction of dinitrosyl-iron(II)-L-cysteine with vascular tissue. Dinitrosyl-iron(II)-L-cysteine was a potent activator of purified soluble guanylyl cyclase (EC50 10 nM with and 100 nM without superoxide dismutase) and relaxed noradrenaline-precontracted segments of endothelium-denuded rabbit femoral artery (EC50 10 nM superoxide dismutase). Pre-incubation (5 min; 310 K) of endothelium-denuded rabbit aortic segments with dinitrosyl-iron(II)-L-cysteine (0.036-3.6 mM) resulted in a concentration-dependent formation of a dinitrosyl-iron(II) complex with protein thiol groups, as detected by ESR spectroscopy. While the complex with proteins was stable for 2 h at 310 K, dinitrosyl-iron(II)-L-cysteine in aqueous solution (36-360 microM) decomposed completely within 15 min, as indicated by disappearance of its isotropic ESR signal at gav = 2.03 (293 K). Aortic segments pre-incubated with dinitrosyl-iron(II)-L-cysteine released a labile vasodilating and guanylyl cyclase activating factor. Perfusion of these segments with N-acetyl-L-cysteine resulted in the generation of a low molecular weight dinitrosyl-iron(II)-dithiolate from the dinitrosyl-iron(II) complex with proteins, as revealed by the shape change of the ESR signal at 293 K. The low molecular weight dinitrosyl-iron(II)-dithiolate accounted for an enhanced guanylyl cyclase activation and vasodilation induced by the aortic effluent. We conclude that nitric oxide (NO) produced by, or acting on vascular cells can be stabilized and stored as a dinitrosyl-iron(II) complex with protein thiols, and can be released from cells in the form of a low molecular weight dinitrosyl-iron(II)-dithiolate by intra- and extracellular thiols.
Mülsch
A
A
Department of Applied Physiology, University of Freiburg, Germany.
Mordvintcev
P
P
Vanin
A F
AF
Busse
R
R
D000818:Animals; D001011:Aorta / Q000378:metabolism; D001808:Blood Vessels / Q000187:drug effects / Q000378:metabolism*; D003545:Cysteine / Q000031:analogs & derivatives* / Q000494:pharmacology; D004578:Electron Spin Resonance Spectroscopy; D004789:Enzyme Activation / Q000187:drug effects; D005263:Femoral Artery / Q000378:metabolism; D006162:Guanylate Cyclase / Q000378:metabolism*; D009569:Nitric Oxide / Q000494:pharmacology*; D009589:Nitrogen Oxides / Q000494:pharmacology*; D011485:Protein Binding; D011506:Proteins / Q000378:metabolism*; D011817:Rabbits; D013438:Sulfhydryl Compounds / Q000494:pharmacology*; D014664:Vasodilation / Q000187:drug effects*
D009589:Nitrogen Oxides; D011506:Proteins; D013438:Sulfhydryl Compounds; C072081:dinitrosyl-iron(II)-cysteine; D009569:Nitric Oxide; D006162:Guanylate Cyclase; D003545:Cysteine