Advertisement

Amino Acids

, Volume 48, Issue 7, pp 1695–1706 | Cite as

Carbonic anhydrases are producers of S-nitrosothiols from inorganic nitrite and modulators of soluble guanylyl cyclase in human platelets

  • Erik Hanff
  • Anke Böhmer
  • Maximilian Zinke
  • Stepan Gambaryan
  • Alexandra Schwarz
  • Claudiu T. Supuran
  • Dimitrios TsikasEmail author
Original Article

Abstract

Nitric oxide (NO), S-nitrosoglutathione (GSNO) and S-nitrosocysteine are highly potent signaling molecules, acting both by cGMP-dependent and cGMP-independent mechanisms. The NO metabolite nitrite (NO2 ) is a major NO reservoir. Hemoglobin, xanthine oxidoreductase and carbonic anhydrase (CA) have been reported to reduce/convert nitrite to NO. We evaluated the role and the physiological importance of CA for an extra-platelet CA/nitrite/NO/cGMP pathway in human platelets. Authentic NO was analyzed by an NO-sensitive electrode. GSNO and GS15NO were measured by liquid chromatography–tandem mass spectrometry (LC–MS/MS). cGMP was determined by LC–MS/MS or RIA. In reduced glutathione (GSH) containing aqueous buffer (pH 7.4), human and bovine erythrocytic CAII-mediated formation of GSNO from nitrite and GS15NO from 15N-nitrite. In the presence of l-cysteine and GSH, this reaction was accompanied by NO release. Incubation of nitrite with bovine erythrocytic CAII and recombinant soluble guanylyl cyclase resulted in cGMP formation. Upon incubation of nitrite with bovine erythrocytic CAII and washed human platelets, cGMP and P-VASPS239 were formed in the platelets. This study provides the first evidence that extra-platelet nitrite and erythrocytic CAII may modulate platelet function in a cGMP-dependent manner. The new nitrite-dependent CA activity may be a general principle and explain the cardioprotective effects of inorganic nitrite in the vasculature. We propose that nitrous acid (ONOH) is the primary CA-catalyzed reaction product of nitrite.

Keywords

Carbonic anhydrase cGMP Nitric oxide Nitrite S-Nitrosothiols Platelets 

Abbreviations

AlbSNO

S-Nitrosoalbumin

BSA

Bovine serum albumin

CA

Carbonic anhydrase

cGMP

Cyclic guanosine monophosphate

CVD

Cardiovascular disease

CysSH

Reduced l-cysteine

CysSNO

S-Nitrosocysteine

GC

Gas chromatography

GSH

Glutathione

GSNO

S-Nitrosoglutathione

GTP

Guanosine triphosphate

HbSNO

S-Nitrosohemoglobin

HMM

High-molecular-mass

IS

Internal standard

LC

Liquid chromatography

LC–MS/MS

Liquid chromatography–tandem mass spectrometry

LMM

Low-molecular-mass

NO

Nitric oxide

NR

Nitrate reductase

PKG

Protein kinase

P-VASPS239

VASP phosphorylated at Ser239

RSNO

S-Nitrosothiol

sGC

Soluble guanylyl cyclase

SIDM

Stable-isotope dilution mass spectrometry

SNP

Sodium nitroprusside

VASP

Vasodilator-stimulated phosphoprotein

Notes

Acknowledgments

We are grateful to Prof. R. Seifert from the Institute of Pharmacology and Prof. V. Kaever from the Core Unit Metabolomics, both Hannover Medical School, for providing us with the recombinant soluble guanylyl cyclase and for the LC–MS/MS analysis of cGMP. This work was supported by the Deutsche Forschungsgemeinschaft (DFG) grant TS60/4-1 (to D.T.).

Compliance with ethical standards

Conflicts of interest

The authors declare that they have no conflict of interest.

Ethical statement

The study on human platelets and human erythrocytes was approved by the Ethics Committee of the Hannover Medical School.

References

  1. Aamand R, Dalsgaard T, Jensen FB, Simonsen U, Roepstorff A, Fago A (2009) Generation of nitric oxide from nitrite by carbonic anhydrase: a possible link between metabolic activity and vasodilation. Am J Physiol Heart Circ Physiol 297:H2068–H2074CrossRefPubMedGoogle Scholar
  2. Adamczyk K, Prémont-Schwarz M, Pines D, Pines E, Nibbering ET (2009) Real-time observation of carbonic acid formation in aqueous solution. Science 326:1690–1694CrossRefPubMedGoogle Scholar
  3. Alvarez BV, Quon AL, Mullen J, Casey JR (2013) Quantification of carbonic anhydrase gene expression in ventricle of hypertrophic and failing human heart. BMC Cardiovasc Disord 13:2. doi: 10.1186/1471-2261-13-2 CrossRefPubMedPubMedCentralGoogle Scholar
  4. Beste KY, Burhenne H, Kaever V, Stasch J, Seifert R (2012) Nucleotidyl cyclase activity of soluble guanylyl cyclase α1β1. Biochemistry (NY) 51:194–204CrossRefGoogle Scholar
  5. Böhmer A, Mitschke A, Reib A, Gutzki FM, Tsikas D (2012) 18O-Labeled nitrous acid and nitrite: synthesis, characterization, and oxyhemoglobin-catalyzed oxidation to 18O-labeled nitrate. Anal Biochem 421:770–772CrossRefPubMedGoogle Scholar
  6. Burkhart JM, Vaudel M, Gambaryan S et al (2012) The first comprehensive and quantitative analysis of human platelet protein composition allows the comparative analysis of structural and functional pathways. Blood 120:e73–e822012CrossRefPubMedGoogle Scholar
  7. Chai YC, Jung CH, Lii CK et al (1991) Identification of an abundant S-thiolated rat liver protein as carbonic anhydrase III; characterization of S-thiolation and dethiolation reactions. Arch Biochem Biophys 284:270–278CrossRefPubMedGoogle Scholar
  8. Chobanyan-Jürgens K, Schwarz A, Böhmer A et al (2012) Renal carbonic anhydrases are involved in the reabsorption of endogenous nitrite. Nitric Oxide 26:126–131CrossRefPubMedGoogle Scholar
  9. de Belder AJ, MacAllister R, Radomski MW, Moncada S, Vallance PJ (1994) Effects of S-nitroso-glutathione in the human forearm circulation: evidence for selective inhibition of platelet activation. Cardiovasc Res 28:691–694CrossRefPubMedGoogle Scholar
  10. Gambaryan S, Tsikas D (2015) A review and discussion of platelet nitric oxide and nitric oxide synthase: do blood platelets produce nitric oxide from l-arginine or nitrite. Amino Acids 47:1779–1793CrossRefPubMedGoogle Scholar
  11. Gambaryan S, Kobsar A, Hartmann S et al (2008) NO-synthase-/NO-independent regulation of human and murine platelet soluble guanylyl cyclase activity. J Thromb Haemost 6:1376–1384CrossRefPubMedGoogle Scholar
  12. Gao G, Xuan C, Yang Q, Liu XC, Liu ZG, He GW (2013) Identification of altered plasma proteins by proteomic study in valvular heart diseases and the potential clinical significance. PLoS One 8:e72111CrossRefPubMedPubMedCentralGoogle Scholar
  13. Giustarini D, Milzani A, Dalle-Donne I, Rossi R (2007) Detection of S-nitrosothiols in biological fluids: a comparison among the most widely applied methodologies. J Chromatogr B 851:124–139CrossRefGoogle Scholar
  14. Gladwin MT, Raat NJ, Shiva S et al (2006) Nitrite as a vascular endocrine nitric oxide reservoir that contributes to hypoxic signaling, cytoprotection, and vasodilation. Am J Physiol Heart Circ Physiol 291:H2026–H2035CrossRefPubMedGoogle Scholar
  15. Hanff E, Böhmer A, Jordan J, Tsikas D (2014) Stable-isotope dilution LC–MS/MS measurement of nitrite in human plasma after its conversion to S-nitrosoglutathione. J Chromatogr B 970:44–52CrossRefGoogle Scholar
  16. Ho C, Sturtevant JM (1963) The kinetics of the hydration of carbon dioxide at 25 degrees. J Biol Chem 238:3499–3501PubMedGoogle Scholar
  17. Keimer R, Stutzer FK, Tsikas D, Troost R, Gutzki FM, Frölich JC (2003) Lack of oxidative stress during sustained therapy with isosorbide dinitrate and pentaerythrityl tetranitrate in healthy humans: a randomized, double-blind crossover study. J Cardiovasc Pharmacol 41:284–292CrossRefPubMedGoogle Scholar
  18. Lesnichin SB, Shenderovich IG, Muljati T, Silverman D, Limbach HH (2011) Intrinsic proton-donating power of zinc-bound water in a carbonic anhydrase active site model estimated by NMR. J Am Chem Soc 133:11331–11338CrossRefPubMedPubMedCentralGoogle Scholar
  19. Li S, Whorton AR (2005) Identification of stereoselective transporters for S-nitroso-l-cysteine: role of LAT1 and LAT2 in biological activity of S-nitrosothiols. J Biol Chem 280:20102–20110CrossRefPubMedGoogle Scholar
  20. Lima B, Forrester MT, Hess DT, Stamler JS (2010) S-Nitrosylation in cardiovascular signaling. Circ Res 106:633–646CrossRefPubMedPubMedCentralGoogle Scholar
  21. Liu C, Waijh N, Liu X et al (2015) Mechanisms of human erythrocytic bioactivation of nitrite. J Biol Chem 290:1281–1294CrossRefPubMedGoogle Scholar
  22. MacAllister RJ, Calver AL, Riezebos J, Collier J, Vallance P (1995) Relative potency and arteriovenous selectivity of nitrovasodilators on human blood vessels: an insight into the targeting of nitric oxide delivery. J Pharmacol Exp Ther 273:154–160PubMedGoogle Scholar
  23. Moncada S, Higgs A (1993) The l-arginine-nitric oxide pathway. N Engl J Med 329:2002–2012CrossRefPubMedGoogle Scholar
  24. Monti SM, Supuran CT, De Simone G (2013) Anticancer carbonic anhydrase inhibitors: a patent review (2008–2013). Expert Opin Ther Pat 23:737–749CrossRefPubMedGoogle Scholar
  25. Palmer LA, Doctor A, Chhabra P, Sheram ML, Laubach VE, Karlinsey MZ, Forbes MS, Macdonald T, Gaston B (2007) S-Nitrosothiols signal hypoxia-mimetic vascular pathology. J Clin Invest 117:2592–2601CrossRefPubMedPubMedCentralGoogle Scholar
  26. Pluta RM, Dejam A, Grimes G, Gladwin MT, Oldfield EH (2005) Nitrite infusions to prevent delayed cerebral vasospasm in a primate model of subarachnoid hemorrhage. JAMA 293:1477–1484CrossRefPubMedGoogle Scholar
  27. Sandmann J, Schwedhelm KS, Tsikas D (2005) Specific transport of S-nitrosocysteine in human red blood cells: implications for formation of S-nitrosothiols and transport of NO bioactivity within the vasculature. FEBS Lett 579:4119–4124CrossRefPubMedGoogle Scholar
  28. Schneider JY, Rothmann S, Schröder F, Langen J, Lücke T, Mariotti F, Huneau JF, Frölich JC, Tsikas D (2015) Effects of chronic oral l-arginine administration on the l-arginine/NO pathway in patients with peripheral arterial occlusive disease or coronary artery disease: l-Arginine prevents renal loss of nitrite, the major NO reservoir. Amino Acids 47:1961–1974CrossRefPubMedGoogle Scholar
  29. Tafreshi NK, Lloyd MC, Bui MM, Gillies RJ, Morse DL (2014) Carbonic anhydrase IX as an imaging and therapeutic target for tumors and metastases. Subcell Biochem 75:221–254CrossRefPubMedPubMedCentralGoogle Scholar
  30. Takakura M, Yokomizo A, Tanaka Y, Kobayashi M, Jung G, Banno M, Sakuma T, Imada K, Oda Y, Kamita M, Honda K, Yamada T, Naito S, Ono M (2012) Carbonic anhydrase I as a new plasma biomarker for prostate cancer. ISRN Oncol 2012:768190PubMedPubMedCentralGoogle Scholar
  31. Truppo E, Supuran CT, Sandomenico A et al (2012) Carbonic anhydrase VII is S-glutathionylated without loss of catalytic activity and affinity for sulfonamide inhibitors. Bioorg Med Chem Lett 22:1560–1564CrossRefPubMedGoogle Scholar
  32. Tsikas D, Sandmann J, Rossa S, Gutzki FM, Frölich JC (1999a) Investigations of S-transnitrosylation reactions between low- and high-molecular-weight S-nitroso compounds and their thiols by high-performance liquid chromatography and gas chromatography-mass spectrometry. Anal Biochem 270:231–241CrossRefPubMedGoogle Scholar
  33. Tsikas D, Ikic M, Tewes KS, Raida M, Frölich JC (1999b) Inhibition of platelet aggregation by S-nitroso-cysteine via cGMP-independent mechanisms: evidence of inhibition of thromboxane A2 synthesis in human blood platelets. FEBS Lett 442:162–166CrossRefPubMedGoogle Scholar
  34. Tsikas D, Sandmann J, Denker K, Frölich JC (2000) Is S-nitrosoglutathione formed in nitric oxide synthase incubates? FEBS Lett 483:83–84CrossRefPubMedGoogle Scholar
  35. Tsikas D, Denker K, Frölich JC (2001) Artifactual-free analysis of S-nitrosoglutathione and S-nitroglutathione by neutral-pH, anion-pairing, high-performance liquid chromatography. Study on peroxynitrite-mediated S-nitration of glutathione to S-nitroglutathione under physiological conditions. J Chromatogr A 915:107–116CrossRefPubMedGoogle Scholar
  36. Tsikas D, Sandmann J, Frölich JC (2002) Measurement of S-nitrosoalbumin by gas chromatography-mass spectrometry. III. Quantitative determination in human plasma after specific conversion of the S-nitroso group to nitrite by cysteine and Cu2+ via intermediate formation of S-nitrosocysteine and nitric oxide. J Chromatogr B 772:335–346CrossRefGoogle Scholar
  37. Tsikas D, Schwarz A, Stichtenoth DO (2010) Simultaneous measurement of [15N]nitrate and [15N]nitrite enrichment and concentration in urine by gas chromatography mass spectrometry as pentafluorobenzyl derivatives. Anal Chem 82:2585–2587CrossRefPubMedGoogle Scholar
  38. Tsikas D, Schmidt M, Böhmer A, Zoerner AA, Gutzki FM, Jordan J (2013a) UPLC-MS/MS measurement of S-nitrosoglutathione (GSNO) in human plasma solves the S-nitrosothiol concentration enigma. J Chromatogr B 927:147–157CrossRefGoogle Scholar
  39. Tsikas D, Sutmöller K, Maassen M et al (2013b) Even and carbon dioxide independent distribution of nitrite between plasma and erythrocytes of healthy humans at rest. Nitric Oxide 31:31–37CrossRefPubMedGoogle Scholar
  40. Wang ST, Chen HW, Sheen LY, Li CK (1997) Methionine and cysteine affect glutathione level, glutathione-related enzyme activities and the expression of glutathione S-transferase isozymes in rat hepatocytes. J Nutr 127:2135–2141PubMedGoogle Scholar

Copyright information

© Springer-Verlag Wien 2016

Authors and Affiliations

  • Erik Hanff
    • 1
  • Anke Böhmer
    • 1
  • Maximilian Zinke
    • 1
  • Stepan Gambaryan
    • 2
    • 3
  • Alexandra Schwarz
    • 1
  • Claudiu T. Supuran
    • 4
  • Dimitrios Tsikas
    • 1
    Email author
  1. 1.Centre of Pharmacology and ToxicologyHannover Medical SchoolHannoverGermany
  2. 2.Institute of Evolutionary Physiology and BiochemistryRussian Academy of SciencesSt. PetersburgRussia
  3. 3.Department of Cytology and HistologyS. Petersburg State UniversityS. PetersburgRussia
  4. 4.Dipartimento Neurofarba, Sezione di Scienze FarmaceuticheUniversità degli Studi di FirenzeSesto FiorentinoItaly

Personalised recommendations