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Amino Acids

, Volume 13, Issue 3–4, pp 355–367 | Cite as

Novel findings on the copper catalysed oxidation of cysteine

  • L. Pecci
  • G. Montefoschi
  • G. Musci
  • D. Cavallini
Review Article

Summary

The oxidation of cysteine (RSH) has been studied by using O2, ferricytochrome c (Cyt c) and nitro blue tetrazolium (NBT) as electron acceptors. The addition of 200μM CuII to a solution of 2mM cysteine, pH 7.4, produces an absorbance with a peak at 260 nm and a shoulder at 300 nm. Generation of a cuprous bis-cysteine complex (RS-CuI-SR) is responsible for this absorbance. In the absence of O2 the absorbance is stable for long time while in the presence of air it vanishes slowly only when the cysteine excess is consumed. The neocuproine assay and the EPR analysis show that the metal remains reduced in the course of the oxidation of cysteine returning to the oxidised form at the end of reaction when all RSH has been oxidised to RSSR. Addition of CuII enhances the reduction rate of Cyt c and of NBT by cysteine also under anaerobiosis indicating the occurrence of a direct reduction of the acceptor by the complex. It is concluded that the cuprous bis-cysteine complex (RS-CuI-SR) is the catalytic species involved in the oxidation of cysteine. The novel finding of the stability of the complex together with the metal remaining in the reduced form during the oxidation suggest sulfur as the electron donor in the place of the metal ion.

Keywords

Amino acids Cysteine Copper catalysis Cuprous complex 

Abbreviations

RSH

cysteine

RS

cysteine in the thiolate form

RS·

thiyl radical of cysteine

RSSR

cystine

Cyt c

cytochrome c

SOD

superoxide dismutase

NBT

nitro blue tetrazolium

NBF

nitro blue formazan

DTNB

5,5′-dithiobis-2-nitrobenzoic acid

DTPA

diethylenetriaminepentaacetic acid

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References

  1. Bielski BHJ, Richter HW (1977) A study of the superoxide radical chemistry by stoppedflow radiolysis and radiation induced oxygen consumption. J Am Chem Soc 99: 3019–3023Google Scholar
  2. Bielski BHJ, Allen AO (1977) Mechanism of the disproportionation of superoxide radicals. J Phys Chem 81: 1048–1050Google Scholar
  3. Bielski BHJ, Shiue GG, Bajuk S (1980) Reduction of nitro blue tetrazolium by CO2 , and O2 radicals. J Phys Chem 84: 830–833Google Scholar
  4. Butler J, Koppenol WH, Margoliash E (1982) Kinetics and mechanism of the reduction of ferricytochrome c by the superoxide anion. J Biol Chem 257: 10747–10750PubMedGoogle Scholar
  5. Cavallini D, De Marco C, Dupré S (1968) Luminol chemiluminescence studies of the oxidation of cysteine and other thiols to disulfides. Arch Biochem Biophys 124: 18–26PubMedGoogle Scholar
  6. Cavallini D, De Marco C, Duprè S, Rotilio G (1969) The copper catalysed oxidation of cysteine to cystine. Arch Biochem Biophys 130: 354–361PubMedGoogle Scholar
  7. Davis FJ, Gilbert BC, Norman ROC (1983) Electron spin resonance studies. Characterization of copper(II) complexes in the oxidation of D-penicillamine, L-cysteine and related sulfur-containig compounds. J Chem Soc Perkin Trans 2: 1763–1771Google Scholar
  8. Deters D, Jurgen HJ, Weser U (1994) Transient thiyl radicals in yeast copper(I) thionein. Biochem Biophys Acta 1208: 344–347PubMedGoogle Scholar
  9. Dikalov S, Khramtsov V, Zimmer G (1983) Determination of rate constants of the reactions of thiols with the superoxide radical by electron paramagnetic resonance: critical remarks on spectrophotometric approaches. Arch Biochem Biophys 326: 207–218Google Scholar
  10. Duprè S, Federici G, Santoro L, Rossi Fanelli MR, Cavallini D (1975) The involvement of superoxide anions in the autoxidation of various cofactors of cysteamine oxygenase. Mol Cell Biochem 9: 149–155PubMedGoogle Scholar
  11. Graf L, Fallab S (1964) Zum Reaktionmechanismus von Oxidasen. Experientia 20: 46–47PubMedGoogle Scholar
  12. Hanna PM, Mason RP (1992) Direct evidence for inhibition of free radicals formation from Cu(I) and hydrogen peroxide by glutathione and other potential ligands using the EPR spin-trapping technique. Arch Biochem Biophys 295: 205–213PubMedGoogle Scholar
  13. Harmann LS, Mottley C, Mason RP (1984) Free radical metabolites of L-cysteine oxidation. J Biol Chem 259: 5606–5611PubMedGoogle Scholar
  14. Hoffman MZ, Bayou E (1972) One-electron reduction of the disulfide linkage in aqueous solution. Formation, protonation and decay kinetics of the RSSR radical. J Am Chem Soc 94: 7950–7957Google Scholar
  15. Kolthoff JM, Stricks W (1951) Polarographic investigations of reactions in aqueous solutions containing copper and cysteine (cystine). J Am Chem Soc 73: 1728–1733Google Scholar
  16. Micic OJ, Nenadovic MT, Carapellucci PA (1978) Solvent participation in reactions. 2. Reactions of cystamine radical. J Am Chem Soc 100: 2209–2212Google Scholar
  17. Misra HP (1974) Generation of superoxide free radical during the autoxidation of thiols. J Biol Chem 249: 2151–2155PubMedGoogle Scholar
  18. Peisach J, Blumber WE (1974) Structural implications derived from the analysis of electron paramagnetic resonance spectra of natural and artificial copper proteins. Arch Biochem Biophys 165: 691–708PubMedGoogle Scholar
  19. Riddles PW, Blakeley RL, Zerner B (1983) Reassesment of Ellman's reagent. Meth Enzymol 91: 49–55PubMedGoogle Scholar
  20. Rossouw SD, Wilken-Jorden TJ (1935) Studies on the origin of sulfur in wool. Biochem J 29: 219–224Google Scholar
  21. Saez G, Thornalley PJ, Hill HAO, Hems R, Bannister JV (1982) The production of free radicals during the autoxidation of cysteine and their effect on isolated rat hepatocytes. Biochim Biophys Acta 719: 24–31PubMedGoogle Scholar
  22. Schafer K, Bonifacic M, Bahneman D, Asmus K-D (1978) Addition of oxygen to organic sulfur radicals. J Phys Chem 82: 2777–2780Google Scholar
  23. Smith GF, McCurdy (1952) 2,9-dimethyl-1,10-phenanthroline. New specific spectrophotometric determination of copper. Anal Chem 24: 371–373Google Scholar
  24. Spear N, Aust SD (1995) Hydroxylation of deoxyguanosine in DNA by copper and thiols. Arch Biochem Biophys 317: 142–148PubMedGoogle Scholar
  25. Suzuki Y, Lyall V, Biber TUL, Ford GD (1990) A modified technique for the measurement of sulfhydryl groups oxidized by reactive oxygen intermediates. Free Rad Biol Med 9: 479–484PubMedGoogle Scholar
  26. Vortisch V, Kroneck P, Hemmerich P (1976) Model studies on the coordination of copper in enzymes. IV. Structure and stability of cuprous complexes with sulfur-containing ligands. J Am Chem Soc 98: 2821–2826Google Scholar
  27. Wefers H, Sies H (1983) Oxidation of glutathione by the superoxide radical to the disulfide and the sulfonate yielding singlet oxygen. Eur J Biochem 137: 29–36PubMedGoogle Scholar
  28. Winterbourn CC, Metodiewa D (1994) The reaction of superoxide with reduced glutathione. Arch Biochem Biophys 314: 284–290PubMedGoogle Scholar
  29. Younes M, Weser U (1977) Superoxide dismutase activity of copper-penicillamine: possible involvement of Cu(I) stabilized sulfur radical. Biochem Biophys Res Comm 78: 1247–1253PubMedGoogle Scholar
  30. Zhao R, Lind J, Merenyi G, Eriksen TR (1994) Kinetics of one-electron oxidation of thiols and hydrogen abstraction by thiyl radicals fromα-amino C-H bonds. J Am Chem Soc 116: 12010–12015Google Scholar

Copyright information

© Springer-Verlag 1997

Authors and Affiliations

  • L. Pecci
    • 1
    • 2
  • G. Montefoschi
    • 1
    • 2
  • G. Musci
    • 3
  • D. Cavallini
    • 1
    • 2
  1. 1.Dipartimento di Scienze Biochimiche “A. Rossi Fanelli”Università di Roma “La Sapienza”RomaItaly
  2. 2.Centro di Biologia Molecolare del CNRUniversità di Roma “La Sapienza”Roma
  3. 3.Dipartimento di Chimica Organica e BiologicaUniversità di MessinaMessinaItaly

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