Skip to main content
SpringerLink
Log in

Novel findings on the copper catalysed oxidation of cysteine

  • Review Article
  • Published:
Amino Acids Aims and scope Submit manuscript

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.

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

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

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

References

  • 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: 3019–3023

    Google Scholar 

  • Bielski BHJ, Allen AO (1977) Mechanism of the disproportionation of superoxide radicals. J Phys Chem 81: 1048–1050

    Google Scholar 

  • Bielski BHJ, Shiue GG, Bajuk S (1980) Reduction of nitro blue tetrazolium by CO2 −, and O2 − radicals. J Phys Chem 84: 830–833

    Google Scholar 

  • Butler J, Koppenol WH, Margoliash E (1982) Kinetics and mechanism of the reduction of ferricytochrome c by the superoxide anion. J Biol Chem 257: 10747–10750

    PubMed  Google Scholar 

  • 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: 18–26

    PubMed  Google Scholar 

  • Cavallini D, De Marco C, Duprè S, Rotilio G (1969) The copper catalysed oxidation of cysteine to cystine. Arch Biochem Biophys 130: 354–361

    PubMed  Google Scholar 

  • 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: 1763–1771

    Google Scholar 

  • Deters D, Jurgen HJ, Weser U (1994) Transient thiyl radicals in yeast copper(I) thionein. Biochem Biophys Acta 1208: 344–347

    PubMed  Google Scholar 

  • 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: 207–218

    Google Scholar 

  • 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: 149–155

    PubMed  Google Scholar 

  • Graf L, Fallab S (1964) Zum Reaktionmechanismus von Oxidasen. Experientia 20: 46–47

    PubMed  Google Scholar 

  • 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: 205–213

    PubMed  Google Scholar 

  • Harmann LS, Mottley C, Mason RP (1984) Free radical metabolites of L-cysteine oxidation. J Biol Chem 259: 5606–5611

    PubMed  Google Scholar 

  • 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: 7950–7957

    Google Scholar 

  • Kolthoff JM, Stricks W (1951) Polarographic investigations of reactions in aqueous solutions containing copper and cysteine (cystine). J Am Chem Soc 73: 1728–1733

    Google Scholar 

  • Micic OJ, Nenadovic MT, Carapellucci PA (1978) Solvent participation in reactions. 2. Reactions of cystamine radical. J Am Chem Soc 100: 2209–2212

    Google Scholar 

  • Misra HP (1974) Generation of superoxide free radical during the autoxidation of thiols. J Biol Chem 249: 2151–2155

    PubMed  Google Scholar 

  • 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: 691–708

    PubMed  Google Scholar 

  • Riddles PW, Blakeley RL, Zerner B (1983) Reassesment of Ellman's reagent. Meth Enzymol 91: 49–55

    PubMed  Google Scholar 

  • Rossouw SD, Wilken-Jorden TJ (1935) Studies on the origin of sulfur in wool. Biochem J 29: 219–224

    Google Scholar 

  • 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: 24–31

    PubMed  Google Scholar 

  • Schafer K, Bonifacic M, Bahneman D, Asmus K-D (1978) Addition of oxygen to organic sulfur radicals. J Phys Chem 82: 2777–2780

    Google Scholar 

  • Smith GF, McCurdy (1952) 2,9-dimethyl-1,10-phenanthroline. New specific spectrophotometric determination of copper. Anal Chem 24: 371–373

    Google Scholar 

  • Spear N, Aust SD (1995) Hydroxylation of deoxyguanosine in DNA by copper and thiols. Arch Biochem Biophys 317: 142–148

    PubMed  Google Scholar 

  • 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: 479–484

    PubMed  Google Scholar 

  • 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: 2821–2826

    Google Scholar 

  • 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: 29–36

    PubMed  Google Scholar 

  • Winterbourn CC, Metodiewa D (1994) The reaction of superoxide with reduced glutathione. Arch Biochem Biophys 314: 284–290

    PubMed  Google Scholar 

  • Younes M, Weser U (1977) Superoxide dismutase activity of copper-penicillamine: possible involvement of Cu(I) stabilized sulfur radical. Biochem Biophys Res Comm 78: 1247–1253

    PubMed  Google Scholar 

  • 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: 12010–12015

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Dipartimento di Scienze Biochimiche “A. Rossi Fanelli”, Università di Roma “La Sapienza”, Piazzale A. Moro 5, I-00185, Roma, Italy

    L. Pecci, G. Montefoschi & D. Cavallini

  2. Centro di Biologia Molecolare del CNR, Università di Roma “La Sapienza”, Roma

    L. Pecci, G. Montefoschi & D. Cavallini

  3. Dipartimento di Chimica Organica e Biologica, Università di Messina, Messina, Italy

    G. Musci

Authors
  1. L. Pecci
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. G. Montefoschi
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. G. Musci
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. D. Cavallini
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

Dedicated to prof. A. Ballio ob the occasion of his 75th birthday.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Pecci, L., Montefoschi, G., Musci, G. et al. Novel findings on the copper catalysed oxidation of cysteine. Amino Acids 13, 355–367 (1997). https://doi.org/10.1007/BF01372599

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01372599

Keywords

Search

Navigation