Search tips
Search criteria 


Logo of jcinvestThe Journal of Clinical InvestigationCurrent IssueArchiveSubscriptionAbout the Journal
J Clin Invest. 1993 April; 91(4): 1582–1589.
PMCID: PMC288134

NO forms an adduct with serum albumin that has endothelium-derived relaxing factor-like properties.


Recent evidence suggests that sulfhydryl species can react with oxides of nitrogen under physiologic conditions and thereby stabilize endothelium-derived relaxing factor (EDRF) activity, but the presence of a specific in vivo thiol carrier for nitric oxide (NO) remains controversial. The single free sulfhydryl of serum albumin is the most abundant thiol species in plasma (approximately 0.5 mM) and is particularly reactive towards NO. To examine the potential role of serum albumin in endogenous nitric oxide metabolism, we synthesized S-nitroso-BSA (S-NO-BSA), a model S-nitroso-protein, and examined its effects on platelet function and coronary and systemic vascular tone in 16 mongrel dogs. Intravenous bolus S-NO-BSA markedly reduced mean arterial pressure in a dose-dependent manner and proved seven and a half-fold less potent than intravenous nitroglycerin and 10-fold less potent than intravenous S-nitroso-cysteine (half-maximal response of 75 nmol/kg compared to 10 and 7.5 nmol/kg, respectively; P < 0.05); when given by intravenous infusion (half-maximal response = 10 nmol/kg per min), however, S-NO-BSA and nitroglycerin were equipotent. Intravenous bolus S-NO-BSA had a greater duration of action than either nitroglycerin or S-nitroso-cysteine and produced marked prolongation of the template bleeding time associated with dose-dependent inhibition of ex vivo platelet aggregation (half-maximal response approximately 70 nmol/kg). Intracoronary S-NO-BSA increased coronary blood flow (mean +/- SEM) less effectively than nitroprusside, acetylcholine, or S-nitroso-cysteine (165% +/- 24% vs. 315% +/- 82%, 483% +/- 55%, or 475% +/- 66%, respectively; P < 0.05) although with much longer duration of action. On a molar basis, S-nitroso-cysteine proved more effective than S-nitroso-BSA, nitroprusside, or acetylcholine as an epicardial coronary vasodilator. Thus, serum albumin reacts with oxides of nitrogen to form a stable S-nitroso-thiol with properties reminiscent of authentic EDRF supporting the view that protein associated thiol may participate in the action and metabolism of EDRF.

Full text

Full text is available as a scanned copy of the original print version. Get a printable copy (PDF file) of the complete article (1.7M), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References.

Images in this article

Click on the image to see a larger version.

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Furchgott RF, Zawadzki JV. The obligatory role of endothelial cells in the relaxation of arterial smooth muscle by acetylcholine. Nature. 1980 Nov 27;288(5789):373–376. [PubMed]
  • Furchgott RF. Role of endothelium in responses of vascular smooth muscle. Circ Res. 1983 Nov;53(5):557–573. [PubMed]
  • Radomski MW, Palmer RM, Moncada S. Comparative pharmacology of endothelium-derived relaxing factor, nitric oxide and prostacyclin in platelets. Br J Pharmacol. 1987 Sep;92(1):181–187. [PMC free article] [PubMed]
  • Palmer RM, Ferrige AG, Moncada S. Nitric oxide release accounts for the biological activity of endothelium-derived relaxing factor. Nature. 1987 Jun 11;327(6122):524–526. [PubMed]
  • Lancaster JR, Jr, Hibbs JB., Jr EPR demonstration of iron-nitrosyl complex formation by cytotoxic activated macrophages. Proc Natl Acad Sci U S A. 1990 Feb;87(3):1223–1227. [PubMed]
  • Saran M, Michel C, Bors W. Reaction of NO with O2-. implications for the action of endothelium-derived relaxing factor (EDRF). Free Radic Res Commun. 1990;10(4-5):221–226. [PubMed]
  • Ignarro LJ, Edwards JC, Gruetter DY, Barry BK, Gruetter CA. Possible involvement of S-nitrosothiols in the activation of guanylate cyclase by nitroso compounds. FEBS Lett. 1980 Feb 11;110(2):275–278. [PubMed]
  • Loscalzo J. N-Acetylcysteine potentiates inhibition of platelet aggregation by nitroglycerin. J Clin Invest. 1985 Aug;76(2):703–708. [PMC free article] [PubMed]
  • Downes MJ, Edwards MW, Elsey TS, Walters CL. Determination of a non-volatile nitrosamine by using denitrosation and a chemiluminescence analyser. Analyst. 1976 Sep;101(1206):742–748. [PubMed]
  • Pryor WA, Lightsey JW. Mechanisms of nitrogen dioxide reactions: initiation of lipid peroxidation and the production of nitrous Acid. Science. 1981 Oct 23;214(4519):435–437. [PubMed]
  • Palmer RM, Ashton DS, Moncada S. Vascular endothelial cells synthesize nitric oxide from L-arginine. Nature. 1988 Jun 16;333(6174):664–666. [PubMed]
  • Buga GM, Gold ME, Wood KS, Chaudhuri G, Ignarro LJ. Endothelium-derived nitric oxide relaxes nonvascular smooth muscle. Eur J Pharmacol. 1989 Feb 14;161(1):61–72. [PubMed]
  • Hoeffner U, Boulanger C, Vanhoutte PM. Proximal and distal dog coronary arteries respond differently to basal EDRF but not to NO. Am J Physiol. 1989 Mar;256(3 Pt 2):H828–H831. [PubMed]
  • Kelm M, Schrader J. Control of coronary vascular tone by nitric oxide. Circ Res. 1990 Jun;66(6):1561–1575. [PubMed]
  • Ignarro LJ. Biological actions and properties of endothelium-derived nitric oxide formed and released from artery and vein. Circ Res. 1989 Jul;65(1):1–21. [PubMed]
  • Cox DA, Vita JA, Treasure CB, Fish RD, Alexander RW, Ganz P, Selwyn AP. Atherosclerosis impairs flow-mediated dilation of coronary arteries in humans. Circulation. 1989 Sep;80(3):458–465. [PubMed]
  • Ignarro LJ, Lippton H, Edwards JC, Baricos WH, Hyman AL, Kadowitz PJ, Gruetter CA. Mechanism of vascular smooth muscle relaxation by organic nitrates, nitrites, nitroprusside and nitric oxide: evidence for the involvement of S-nitrosothiols as active intermediates. J Pharmacol Exp Ther. 1981 Sep;218(3):739–749. [PubMed]
  • Stamler JS, Simon DI, Osborne JA, Mullins ME, Jaraki O, Michel T, Singel DJ, Loscalzo J. S-nitrosylation of proteins with nitric oxide: synthesis and characterization of biologically active compounds. Proc Natl Acad Sci U S A. 1992 Jan 1;89(1):444–448. [PubMed]
  • Bonnett R, Holleyhead R, Johnson BL, Randall EW. Reaction of acidified nitrite solutions with peptide derivatives: evidence for nitrosamine and thionitrite formation from 15N N.m.r. studies. J Chem Soc Perkin 1. 1975;(22):2261–2241. [PubMed]
  • Stamler JS, Jaraki O, Osborne J, Simon DI, Keaney J, Vita J, Singel D, Valeri CR, Loscalzo J. Nitric oxide circulates in mammalian plasma primarily as an S-nitroso adduct of serum albumin. Proc Natl Acad Sci U S A. 1992 Aug 15;89(16):7674–7677. [PubMed]
  • Sandor T, Spears JR. Statistical considerations on the precision of assessing blood vessel diameter in cine coronary angiography. Comput Biomed Res. 1985 Dec;18(6):531–543. [PubMed]
  • Ludmer PL, Selwyn AP, Shook TL, Wayne RR, Mudge GH, Alexander RW, Ganz P. Paradoxical vasoconstriction induced by acetylcholine in atherosclerotic coronary arteries. N Engl J Med. 1986 Oct 23;315(17):1046–1051. [PubMed]
  • Nabel EG, Ganz P, Gordon JB, Alexander RW, Selwyn AP. Dilation of normal and constriction of atherosclerotic coronary arteries caused by the cold pressor test. Circulation. 1988 Jan;77(1):43–52. [PubMed]
  • Gordon JB, Ganz P, Nabel EG, Fish RD, Zebede J, Mudge GH, Alexander RW, Selwyn AP. Atherosclerosis influences the vasomotor response of epicardial coronary arteries to exercise. J Clin Invest. 1989 Jun;83(6):1946–1952. [PMC free article] [PubMed]
  • Vita JA, Treasure CB, Nabel EG, McLenachan JM, Fish RD, Yeung AC, Vekshtein VI, Selwyn AP, Ganz P. Coronary vasomotor response to acetylcholine relates to risk factors for coronary artery disease. Circulation. 1990 Feb;81(2):491–497. [PubMed]
  • Sibley DH, Millar HD, Hartley CJ, Whitlow PL. Subselective measurement of coronary blood flow velocity using a steerable Doppler catheter. J Am Coll Cardiol. 1986 Dec;8(6):1332–1340. [PubMed]
  • Macho P, Vatner SF. Effects of nitroglycerin and nitroprusside on large and small coronary vessels in conscious dogs. Circulation. 1981 Dec;64(6):1101–1107. [PubMed]
  • Mordvintsev PI, Rudneva VG, Vanin AF, Shimkevich LL, Khodorov BI. Ingibiruiushchee vliianie na agregatsiiu trombotsitov dinitrozil'nykh kompleksov zheleza s nizkomolekuliarnymi ligandami. Biokhimiia. 1986 Nov;51(11):1851–1857. [PubMed]
  • Cooke JP, Stamler J, Andon N, Davies PF, McKinley G, Loscalzo J. Flow stimulates endothelial cells to release a nitrovasodilator that is potentiated by reduced thiol. Am J Physiol. 1990 Sep;259(3 Pt 2):H804–H812. [PubMed]
  • Kowaluk EA, Fung HL. Spontaneous liberation of nitric oxide cannot account for in vitro vascular relaxation by S-nitrosothiols. J Pharmacol Exp Ther. 1990 Dec;255(3):1256–1264. [PubMed]
  • Mellion BT, Ignarro LJ, Myers CB, Ohlstein EH, Ballot BA, Hyman AL, Kadowitz PJ. Inhibition of human platelet aggregation by S-nitrosothiols. Heme-dependent activation of soluble guanylate cyclase and stimulation of cyclic GMP accumulation. Mol Pharmacol. 1983 May;23(3):653–664. [PubMed]
  • Stamler J, Mendelsohn ME, Amarante P, Smick D, Andon N, Davies PF, Cooke JP, Loscalzo J. N-acetylcysteine potentiates platelet inhibition by endothelium-derived relaxing factor. Circ Res. 1989 Sep;65(3):789–795. [PubMed]
  • Chong S, Fung HL. Thiol-mediated catalysis of nitroglycerin degradation by serum proteins. Increase in metabolism was not accompanied by S-nitrosothiol production. Drug Metab Dispos. 1990 Jan-Feb;18(1):61–67. [PubMed]
  • Keen JH, Habig WH, Jakoby WB. Mechanism for the several activities of the glutathione S-transferases. J Biol Chem. 1976 Oct 25;251(20):6183–6188. [PubMed]
  • Bennett BM, Kobus SM, Brien JF, Nakatsu K, Marks GS. Requirement for reduced, unliganded hemoprotein for the hemoglobin- and myoglobin-mediated biotransformation of glyceryl trinitrate. J Pharmacol Exp Ther. 1986 May;237(2):629–635. [PubMed]

Articles from The Journal of Clinical Investigation are provided here courtesy of American Society for Clinical Investigation