Skip to main content

Nitrate Reductase Activity of Mitochondrial Aldehyde Dehydrogenase (ALDH-2) as a Redox Sensor for Cardiovascular Oxidative Stress

  • Protocol
  • First Online:
Advanced Protocols in Oxidative Stress II

Part of the book series: Methods in Molecular Biology ((MIMB,volume 594))

Abstract

In 2002, mitochondrial aldehyde dehydrogenase (ALDH-2) was identified as an organic nitrate bioactivating enzyme. This so-called nitrate reductase activity denitrates nitroglycerin (glycerol trinitrate) to its 1,2-glycerol dinitrate metabolite and nitrite. This reaction relies on reduced thiols at the active site of the enzyme and on the presence of reduced dithiols as the electron source. During bioconversion of nitroglycerin, and also in the presence of reactive oxygen and nitrogen species, the active site thiols of ALDH-2 are oxidized and the enzyme looses its activity. We, therefore, speculated that ALDH-2 activity could be a useful marker for cardiovascular oxidative stress. Indeed, this hypothesis was supported by a number of studies, indicating that ALDH-2 activity is impaired in experimental animal models of increased oxidative stress and may be used for detection of an imbalance of mitochondrial and cellular redox state.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 84.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 159.00
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 109.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

Similar content being viewed by others

References

  1. Chen Z, Zhang J, Stamler JS (2002) Identification of the enzymatic mechanism of nitroglycerin bioactivation. Proc Natl Acad Sci U S A 99:8306–11

    Article  PubMed  CAS  Google Scholar 

  2. Sydow K, Daiber A, Oelze M, Chen Z, August M, Wendt M, Ullrich V, Mulsch A, Schulz E, Keaney JF Jr, Stamler JS, Munzel T (2004) Central role of mitochondrial aldehyde dehydrogenase and reactive oxygen species in nitroglycerin tolerance and cross-tolerance. J Clin Invest 113:482–9

    PubMed  CAS  Google Scholar 

  3. Wenzel P, Hink U, Oelze M, Schuppan S, Schaeuble K, Schildknecht S, Ho KK, Weiner H, Bachschmid M, Munzel T, Daiber A (2007) Role of reduced lipoic acid in the redox regulation of mitochondrial aldehyde dehydrogenase (ALDH-2) activity: Implications for mitochondrial Oxidative stress and nitrate tolerance. J Biol Chem 282:792–9

    Article  PubMed  CAS  Google Scholar 

  4. Chen Z, Stamler JS (2006) Bioactivation of nitroglycerin by the mitochondrial aldehyde dehydrogenase. Trends Cardiovasc Med 16:259–65

    Article  PubMed  CAS  Google Scholar 

  5. Daiber A, Oelze M, Coldewey M, Bachschmid M, Wenzel P, Sydow K, Wendt M, Kleschyov AL, Stalleicken D, Ullrich V, Mulsch A, Munzel T (2004) Oxidative stress and mitochondrial aldehyde dehydrogenase activity: a comparison of pentaerythritol tetranitrate with other organic nitrates. Mol Pharmacol 66:1372–82

    Article  PubMed  CAS  Google Scholar 

  6. Daiber A, Bachschmid M (2007) Enzyme inhibition by peroxynitrite-mediated tyrosine nitration and thiol oxidation. Curr Enzyme Inhib 3:103–17

    Article  CAS  Google Scholar 

  7. Daiber A, Oelze M, Coldewey M, Kaiser K, Huth C, Schildknecht S, Bachschmid M, Nazirisadeh Y, Ullrich V, Mulsch A, Munzel T, Tsilimingas N (2005) Hydralazine is a powerful inhibitor of peroxynitrite formation as a possible explanation for its beneficial effects on prognosis in patients with congestive heart failure. Biochem Biophys Res Commun 338:1865–74

    Article  PubMed  CAS  Google Scholar 

  8. Hink U, Daiber A, Kayhan N, Trischler J, Kraatz C, Oelze M, Mollnau H, Wenzel P, Vahl CF, Ho KK, Weiner H, Munzel T (2007) Oxidative inhibition of the mitochondrial aldehyde dehydrogenase promotes nitroglycerin tolerance in human blood vessels. J Am Coll Cardiol 50:2226–32

    Article  PubMed  CAS  Google Scholar 

  9. Wenzel P, Oelze M, Coldewey M, Hortmann M, Seeling A, Hink U, Mollnau H, Stalleicken D, Weiner H, Lehmann J, Li H, Forstermann U, Munzel T, Daiber A (2007) Heme oxygenase-1: a novel key player in the development of tolerance in response to organic nitrates. Arterioscler Thromb Vasc Biol 27:1729–35

    Article  PubMed  CAS  Google Scholar 

  10. Daiber A, Oelze M, Sulyok S, Coldewey M, Schulz E, Treiber N, Hink U, Mulsch A, Scharffetter-Kochanek K, Munzel T (2005) Heterozygous deficiency of manganese superoxide dismutase in mice (Mn-SOD+/−): A novel approach to assess the role of oxidative stress for the development of nitrate tolerance. Mol Pharmacol 68:579–88

    PubMed  CAS  Google Scholar 

  11. Wenzel P, Muller J, Zurmeyer S, Schuhmacher S, Schulz E, Oelze M, Pautz A, Kawamoto T, Wojnowski L, Kleinert H, Munzel T, Daiber A (2008) ALDH-2 deficiency increases cardiovascular oxidative stress – Evidence for indirect antioxidative properties. Biochem Biophys Res Commun 367:137–143

    Google Scholar 

  12. Szocs K, Lassegue B, Wenzel P, Wendt M, Daiber A, Oelze M, Meinertz T, Munzel T, Baldus S (2007) Increased superoxide production in nitrate tolerance is associated with NAD(P)H oxidase and aldehyde dehydrogenase 2 downregulation. J Mol Cell Cardiol 42:1111–8

    Article  PubMed  Google Scholar 

  13. Saha A, Goldstein S, Cabelli D, Czapski G (1998) Determination of optimal conditions for synthesis of peroxynitrite by mixing acidified hydrogen peroxide with nitrite. Free Radic Biol Med 24:653–9

    Article  PubMed  CAS  Google Scholar 

  14. Wenzel P, Hink U, Oelze M, Seeling A, Isse T, Bruns K, Steinhoff L, Brandt M, Kleschyov AL, Schulz E, Lange K, Weiner H, Lehmann J, Lackner KJ, Kawamoto T, Munzel T, Daiber A (2007) Number of nitrate groups determines reactivity and potency of organic nitrates: a proof of concept study in ALDH-2−/− mice. Brit J Pharmacol 150:526–33

    Article  CAS  Google Scholar 

  15. Gerardin A, Gaudry D, Wantiez D (1982) Gas chromatographic mass spectrometric determination of 1, 2, 3-propanetrioltrinitrate (nitroglycerin) in human plasma using the nitrogen-15 labelled compound as internal standard. Biomed Mass Spectrom 9:333–5

    Article  PubMed  CAS  Google Scholar 

  16. Brien JF, McLaughlin BE, Breedon TH, Bennett BM, Nakatsu K, Marks GS (1986) Biotransformation of glyceryl trinitrate occurs concurrently with relaxation of rabbit aorta. J Pharmacol Exp Ther 237:608–14

    PubMed  CAS  Google Scholar 

  17. Seeling A, Lehmann J (2006) NO-donors, part X (1) : investigations on the stability of pentaerythrityl tetranitrate (PETN) by HPLC-chemoluminescence-N-detection (CLND) versus UV-detection in HPLC. J Pharm Biomed Anal 40:1131–6

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

We thank the German Research Foundation for continuous funding of our ongoing research on nitrate tolerance (SFB 553-C17 to T.M. and A.D.) and the University Hospital Mainz for financial support (MAIFOR to A.D.).

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Andreas Daiber .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2010 Humana Press, a part of Springer Science+Business Media, LLC

About this protocol

Cite this protocol

Daiber, A., Münzel, T. (2010). Nitrate Reductase Activity of Mitochondrial Aldehyde Dehydrogenase (ALDH-2) as a Redox Sensor for Cardiovascular Oxidative Stress. In: Armstrong, D. (eds) Advanced Protocols in Oxidative Stress II. Methods in Molecular Biology, vol 594. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60761-411-1_3

Download citation

  • DOI: https://doi.org/10.1007/978-1-60761-411-1_3

  • Published:

  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-60761-410-4

  • Online ISBN: 978-1-60761-411-1

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics