Analysis of Neuroketal Protein Adducts by Liquid Chromatography-Electrospray Ionization/Tandem Mass Spectrometry

  • Nathalie Bernoud-Hubac
  • Sean S. Davies
  • Olivier Boutaud
  • L. Jackson RobertsII
Part of the Methods in Pharmacology and Toxicology book series (MIPT)


A role of free radicals has been implicated in the pathogenesis of a number of neurological disorders, including amyotrophic lateral sclerosis, Huntington’s disease (HD), Parkinson’s disease (PD), and Alzheimer’s disease (AD) (1, 2, 3, 4, 5, 6, 7, 8, 9). Furthermore, reactive aldehydes produced as products of lipid peroxidation are thought to be key mediators of oxidant injury because of their capacity to covalently modify proteins and DNA (10), and evidence suggests they may be involved in the pathogenesis of neurodegenerative diseases (11, 12, 13). Aggregated cross-linked proteins are characteristic features of neurodegenerative diseases (14). Isoprostanes (IsoPs) are prostaglandin-like compounds formed by free radical-induced peroxidation of arachidonic acid (15). We recently identified the formation of highly reactive γ-ketoaldehydes, now termed isoketals (IsoKs), as products of the IsoP pathway (16). IsoKs are orders of magnitude more reactive than other known reactive products of lipid peroxidation and exhibit a unique proclivity to cross-link proteins (17). Neuroprostanes (NPs) are IsoP-like compounds formed from oxidation of docosahexaenoic acid (DHA) (5), which is highly enriched in the brain (18,19), and levels of neuroprostanes in cerebrospinal fluid (CFS) have been found to be increased in patients with AD (5). Analogous to the formation of IsoKs via the isoprostane pathway, we recently identified the formation of another class of highly reactive γ-ketoaldehydes as products of the neuroprostane pathway, termed neuroketals (NKs) (20).


Amyotrophic Lateral Sclerosis Select Reaction Monitoring Unique Proclivity Protein Adduct Electrospray Ionization Tandem Mass Spectrometry 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


  1. 1.
    Markesbery, W. R. (1997) Oxidative stress hypothesis in Alzheimer’s disease. Free Radic. Biol. Med. 23, 134–147.PubMedCrossRefGoogle Scholar
  2. 2.
    Markesbery, W. R. (1999) The role of oxidative stress in Alzheimer disease. Arch. Neurol. 56, 1449–1452.PubMedCrossRefGoogle Scholar
  3. 3.
    Simonian, N. A. and Coyle, J. T. (1996) Oxidative stress in neurodegenerative diseases. Annu. Rev. Pharmacol. Toxicol. 36, 83–106.PubMedCrossRefGoogle Scholar
  4. 4.
    Perry, G., Nunomura, A., Hirai, K., Takeda, A., Aliev, G., and Smith, M. A. (2000) Oxidative damage in Alzheimer’s disease: the metabolic dimension. Int. J. Dev. Neurosci. 18, 417–421.PubMedCrossRefGoogle Scholar
  5. 5.
    Roberts, L. J., 2nd, Montine, T. J., Markesbery, W. R., Tapper, A. R., Hardy, P., Chemtob, S., et al. (1998) Formation of isoprostane-like compounds (neuroprostanes) in vivo from docosahexaenoic acid. J. Biol. Chem. 273, 13605–13612.PubMedCrossRefGoogle Scholar
  6. 6.
    Montine, T. J., Markesbery, W. R., Morrow, J. D., and Roberts, L. J. (1998) Cerebrospinal fluid F2-isoprostane levels are increased in Alzheimer’s disease. Ann. Neurol. 44, 410–413.PubMedCrossRefGoogle Scholar
  7. 7.
    Montine, T. J., Beal, M. F., Cudkowicz, M. E., O’Donnell, H., Margolin, R. A., McFarland, L., et al. (1999) Increased CSF F2-isoprostane concentration in probable AD. Neurology 52, 562–565.PubMedGoogle Scholar
  8. 8.
    Montine, T. J., Beal, M. F., Robertson, D., Cudkowicz, M. E., Biaggioni, I., O’Donnell, H., et al. (1999) Cerebrospinal fluid F2-isoprostanes are elevated in Huntington’s disease. Neurology 52, 1104–1105.PubMedGoogle Scholar
  9. 9.
    Reich, E. E., Markesbery, W. R., Roberts, L. J., Swift, L. L., Morrow, J. D., and Montine, T. J. (2001) Brain regional quantification of F-ring and D-/E-ring isoprostanes and neuroprostanes in Alzheimer’s disease. Am. J. Pathol. 158, 293–297.PubMedCrossRefGoogle Scholar
  10. 10.
    Esterbauer, H., Schaur, R. J., and Zollner, H. (1991) Chemistry and biochemistry of 4-hydroxynonenal, malonaldehyde and related aldehydes. Free Radic. Biol. Med. 11, 81–128.PubMedCrossRefGoogle Scholar
  11. 11.
    Smith, R. G., Henry, Y. K., Mattson, M. P., and Appel, S. H. (1998) Presence of 4-hydroxynonenal in cerebrospinal fluid of patients with sporadic amyotrophic lateral sclerosis. Ann. Neurol. 44, 696–699.PubMedCrossRefGoogle Scholar
  12. 12.
    Lovell, M. A., Ehmann, W. D., Mattson, M. P., and Markesbery, W. R. (1997) Elevated 4-hydroxynonenal in ventricular fluid in Alzheimer’s disease. Neurobiol. Aging 18, 457–461.PubMedCrossRefGoogle Scholar
  13. 13.
    Montine, K. S., Reich, E., Neely, M. D., Sidell, K. R., Olson, S. J., Markesbery, W. R., and Montine, T. J. (1998) Distribution of reducible 4-hydroxynonenal adduct immunoreactivity in Alzheimer disease is associated with APOE genotype. J. Neuropathol. Exp. Neurol. 57, 415–425.PubMedCrossRefGoogle Scholar
  14. 14.
    Kaytor, M. D., Warren, S. T. (1999) Aberrant protein deposition and neurological disease. J. Biol. Chem. 274, 37,507–37,510.PubMedCrossRefGoogle Scholar
  15. 15.
    Morrow, J. D., Hill, K. E., Burk, R. F., Nammour, T. M., Badr, K. F., and Roberts, L. J., 2nd (1990) A series of prostaglandin F2-like compounds are produced in vivo in humans by a non-cyclooxygenase, free radical-catalyzed mechanism. Proc. Natl. Acad. Sci. USA 87, 9383–9387.PubMedCrossRefGoogle Scholar
  16. 16.
    Brame, C. J., Salomon, R. G., Morrow, J. D., and Roberts, L. J., 2nd (1999) Identification of extremely reactive gamma-ketoaldehydes (isolevuglandins) as products of the isoprostane pathway and characterization of their lysyl protein adducts. J. Biol. Chem. 274, 13,139–13,146.PubMedCrossRefGoogle Scholar
  17. 17.
    Iyer, R. S., Ghosh, S., and Salomon, R. G. (1989) Levuglandin E2 crosslinks proteins. Prostaglandins 37, 471–480.PubMedCrossRefGoogle Scholar
  18. 18.
    Salem, N., Jr., Kim, H.-Y., and Yergery, J. A. (1986) Docosahexaenoic acid∶Membrane function and metaolism, in Health Effects of Polyunsaturated Fatty Acids in Seafoods (Simopoulos. A. P., Kifer, R. R., and Martin, R. E., eds.), Academic Press, Orlando, FL, pp. 263–317.Google Scholar
  19. 19.
    Skinner, E. R., Watt, C., Besson, J. A., and Best, P. V. (1993) Differences in the fatty acid composition of the grey and white matter of different regions of the brains of patients with Alzheimer’s disease and control subjects. Brain 116, 717–725.PubMedCrossRefGoogle Scholar
  20. 20.
    Bernoud-Hubac, N., Davies, S. S., Boutaud, O., Montine, T. J., and Roberts, L. J., 2nd (2001) Formation of highly reactive gamma-ketoaldehydes (neuroketals) as products of the neuroprostane pathway. J. Biol. Chem. 276, 30,964–30,970.PubMedCrossRefGoogle Scholar
  21. 21.
    Taber, D. F., Morrow, J. D., and Roberts, L. J., 2nd (1997) A nomenclature system for the isoprostanes. Prostaglandins 53, 63–67.PubMedCrossRefGoogle Scholar

Copyright information

© Humana Press Inc.,Totowa, NJ 2003

Authors and Affiliations

  • Nathalie Bernoud-Hubac
    • 1
  • Sean S. Davies
    • 1
  • Olivier Boutaud
    • 1
  • L. Jackson RobertsII
    • 1
  1. 1.Departments of Pharmacology and MedicineVanderbilt UniversityNashville

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