Skip to main content
SpringerLink
Log in

Spectroscopic characterization of the [Fe(His)4(Cys)] site in 2Fe-superoxide reductase from Desulfovibrio vulgaris

  • Original Article
  • Published:
JBIC Journal of Biological Inorganic Chemistry Aims and scope Submit manuscript

Abstract

The electronic and vibrational properties of the [Fe(His)4(Cys)] site (Center II) responsible for catalysis of superoxide reduction in the two-iron superoxide reductase (2Fe-SOR) from Desulfovibrio vulgaris have been investigated using the combination of EPR, resonance Raman, UV/visible/near-IR absorption, CD, and VTMCD spectroscopies. Deconvolution of the spectral contributions of Center II from those of the [Fe(Cys)4] site (Center I) has been achieved by parallel investigations of the C13S variant, which does not contain Center I. The resonance Raman spectrum of ferric Center II has been assigned based on isotope shifts for 34S and 15N globally labeled proteins. As for the [Fe(His)4(Cys)] active site in 1Fe-SOR from Pyrococcus furiosus, the spectroscopic properties of ferric and ferrous Center II in D. vulgaris 2Fe-SOR are indicative of distorted octahedral and square-pyramidal coordination geometries, respectively. Differences in the properties of the ferric [Fe(His)4(Cys)] sites in 1Fe- and 2Fe-SORs are apparent in the rhombicity of the S=5/2 ground state (E/D=0.06 and 0.28 in 1Fe- and 2Fe-SORs, respectively), the energy of the CysS−(pπ)→Fe3+(dπ) CT transition (15,150±150 cm−1 and 15,600±150 cm−1 in 1Fe- and 2Fe-SORs, respectively) and in changes in the Fe–S stretching region of the resonance Raman spectrum indicative of a weaker Fe–S(Cys) bond in 2Fe-SORs. These differences are interpreted in terms of small structural perturbations in the Fe coordination sphere with changes in the Fe–S(Cys) bond strength resulting from differences in the peptide N–H...S(Cys) hydrogen bonding within a tetrapeptide bidentate "chelate". Observation of the characteristic intervalence charge transfer transition of a cyano-bridged [FeIII–NC–FeII(CN)5] unit in the near-IR VTMCD spectra of ferricyanide-oxidized samples of both P. furiosus 1Fe-SOR and D. vulgaris 2Fe-SOR has confirmed the existence of novel ferrocyanide adducts of the ferric [Fe(His)4(Cys)] sites in both 1Fe- and 2Fe-SORs.

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

Fig. 1.
Fig. 2.
Fig. 3.
Fig. 4.
Fig. 5.
Fig. 6.
Fig. 7.
Fig. 8.
Fig. 9.
Fig. 10.

Similar content being viewed by others

References

  1. McCord JM (2000) Am J Med 108:652–659

    CAS  PubMed  Google Scholar 

  2. Fridovich I (1998) J Exp Biol 201:1203–1209

    CAS  PubMed  Google Scholar 

  3. Pianzzola MJ, Soubes M, Touati D (1996) J Bacteriol 178:6736–6742

    CAS  PubMed  Google Scholar 

  4. Jenney FE Jr, Verhagen MFJM, Cui X, Adams MWW (1999) Science 286:306–309

    CAS  PubMed  Google Scholar 

  5. Lombard M, Fontecave M, Touati D, Nivière V (2000) J Biol Chem 275:115–121

    CAS  PubMed  Google Scholar 

  6. Adams MWW, Jenney FE Jr, Clay MD, Johnson MK (2002) J Biol Inorg Chem 7:647–652

    Article  CAS  PubMed  Google Scholar 

  7. Coulter ED, Kurtz DM Jr (2002) J Biol Inorg Chem 7:653–658

    Article  PubMed  Google Scholar 

  8. Auchere F, Rusnak F (2002) J Biol Inorg Chem 7:664–667

    Article  CAS  PubMed  Google Scholar 

  9. Imlay JA (2002) J Biol Inorg Chem 7:659–663

    Article  CAS  PubMed  Google Scholar 

  10. Coulter ED, Kurtz DM Jr (2001) Arch Biochem Biophys 394:76–86

    CAS  PubMed  Google Scholar 

  11. Ascenso C, Rusnak FM, Cabrito I, Lima MJ, Naylor S, Moura I, Moura JJG (2000) J Biol Inorg Chem 5:720–729

    CAS  PubMed  Google Scholar 

  12. Abreu IA, Saraiva LM, Carita J, Huber H, Stetter KO, Cabelli DE, Teixeira M (2000) Mol Microbiol 38:322–334

    CAS  PubMed  Google Scholar 

  13. Emerson JP, Coulter ED, Cabelli DE, Phillips RS, Kurtz DM Jr (2002) Biochemistry 41:4348–4357

    Article  CAS  PubMed  Google Scholar 

  14. Coulter ED, Emerson JP, Kurtz DM Jr, Cabelli DE (2000) J Am Chem Soc 122:11555–11556

    CAS  Google Scholar 

  15. Abreu IA, Saraiva LM, Soares CM, Teixeira M, Cabelli DE (2001) J Biol Chem 276:38995–39001

    CAS  PubMed  Google Scholar 

  16. Lombard M, Houée-Levin C, Touati D, Fontecave M, Nivière V (2001) Biochemistry 40:5032–5040

    CAS  PubMed  Google Scholar 

  17. Nivière V, Lombard M, Fontecave M, Houée-Levin C (2001) FEBS Lett 497:171–173

    CAS  PubMed  Google Scholar 

  18. Yeh AP, Hu Y, Jenney FE Jr, Adams MWW, Rees DC (2000) Biochemistry 39:2499–2508

    CAS  PubMed  Google Scholar 

  19. Coelho AV, Matias P, Fülõp V, Thompson A, Gonzalez A, Carrondo MA (1997) J Biol Inorg Chem 2:680–689

    CAS  Google Scholar 

  20. Silaghi-Dumitrescu R, Silaghi-Dumitrescu I, Coulter ED, Kurtz DM Jr (2003) Inorg Chem 42:446–456

    Google Scholar 

  21. Verhagen MFJM, Voorhorst WGB, Kolkman JA, Wolbert RBG, Hagen WR (1993) FEBS Lett 336:13–18

    CAS  PubMed  Google Scholar 

  22. Tavares P, Ravi N, Moura JJG, LeGall J, Huang Y-H, Crouse BR, Johnson MK, Huynh BH, Moura I (1994) J Biol Chem 269:10504–10510

    CAS  PubMed  Google Scholar 

  23. Moura I, Tavares P, Moura JJG, Ravi N, Huynh BH, Liu M-Y, LeGall J (1990) J Biol Chem 265:21596–21602

    CAS  PubMed  Google Scholar 

  24. Clay MD, Jenney FE Jr, Hagedoorn PL, George GN, Adams MWW, Johnson MK (2002) J Am Chem Soc 124:788–805

    CAS  PubMed  Google Scholar 

  25. Clay MD, Jenney FE Jr, Noh HJ, Hagedoorn PL, Adams MWW, Johnson MK (2002) Biochemistry 41:9833–9841

    Article  CAS  PubMed  Google Scholar 

  26. Berthomieu C, Dupeyrat F, Fontecave M, Verméglio A, Nivière V (2002) Biochemistry 41:10360–10368

    Article  CAS  PubMed  Google Scholar 

  27. Mathé C, Mattioli TA, Horner O, Lombard M, Latour J-M, Fontecave M, Nivière V (2002) J Am Chem Soc 124:4966–4967

    Article  PubMed  Google Scholar 

  28. Clay MD, Cosper CA, Jenney FE Jr, Adams MWW, Johnson MK (2003) Proc Natl Acad Sci USA 100:3796–3801

    Article  CAS  PubMed  Google Scholar 

  29. Kurtz DM Jr, Coulter ED (2001) Chemtracts Inorg Chem 14:407–435

    CAS  Google Scholar 

  30. Moura I, Huynh BH, Hausinger RP, LeGall J, Xavier AV, Munck E (1980) J Biol Chem 255:2493–2498

    CAS  PubMed  Google Scholar 

  31. Chen L, Sharma P, LeGall J, Mariano AM, Teixeira M, Xavier AV (1994) Eur J Biochem 226:613–618

    CAS  PubMed  Google Scholar 

  32. Emerson JP, Cabelli DE, Kurtz DM Jr (2003) Proc Natl Acad Sci USA 100:3802–3807

    Article  CAS  PubMed  Google Scholar 

  33. Drozdzewski PM, Johnson MK (1988) Appl Spectrosc 42:1575–1577

    CAS  Google Scholar 

  34. Shelnutt JA, Rousseau DL, Dethemers JK, Margoliash E (1979) Proc Natl Acad Sci USA 76:3865–3871

    CAS  PubMed  Google Scholar 

  35. Johnson MK (1988) In: Que L (ed) Metal clusters in proteins. American Chemical Society, Washington, pp 326–342

  36. Thomson AJ, Cheesman MR, George SJ (1993) Methods Enzymol 226:199–232

    CAS  PubMed  Google Scholar 

  37. Eaton WA, Lovenberg W (1973) In: Lovenberg W (ed) Iron-sulfur proteins. Academic Press, New York, pp 131–162

  38. Jovanovi'c T, Ascenso C, Hazlett KRO, Sikkink R, Krebs C, Litwiller LM, Benson LM, Moura I, Moura JJG, Radolf JD, Huynh BH, Naylor S, Rusnak F (2000) J Biol Chem 275:28439–28448

    CAS  PubMed  Google Scholar 

  39. Solomon EI, Brunold TC, Davis MI, Kemsley JN, Lee S-K, Lehnert N, Neese F, Skulan AJ, Yang T-S, Zhou J (2000) Chem Rev 100:235–349

    CAS  PubMed  Google Scholar 

  40. Oganesyan VS, George SJ, Cheesman MR, Thomson AJ (1999) J Chem Phys 110:762–777

    Article  CAS  Google Scholar 

  41. Gebhard MS, Koch SA, Millar MM, Devlin FJ, Stephens PJ, Solomon EI (1991) J Am Chem Soc 113:1640–1649

    CAS  Google Scholar 

  42. Johnson MK, Robinson AE, Thomson AJ (1982) In: Spiro TG (ed) Iron-sulfur proteins. Wiley-Interscience, New York, pp 367–406

  43. Gebhard MS, Deaton JC, Koch SA, Millar MM, Solomon EI (1990) J Am Chem Soc 112:2217–2231

    CAS  Google Scholar 

  44. Robin MB (1962) Inorg Chem 1:337–342

    CAS  Google Scholar 

  45. Loehr TM (1992) J Raman Spectrosc 23:531–537

    CAS  Google Scholar 

  46. Spiro TG, Czernuszewicz RS (1995) Methods Enzymol 246:416–460

    CAS  PubMed  Google Scholar 

  47. Andrew CR, Sanders-Loehr J (1996) Acc Chem Res 29:365–372

    CAS  Google Scholar 

  48. Andrew CR, Han J, den Blaauwen T, van Pouderoyen G, Vijgenboom E, Canters GW, Loehr TM, Sanders-Loehr J (1997) J Biol Inorg Chem 2:98–107

    CAS  Google Scholar 

  49. Dong S, Spiro TG (1998) J Am Chem Soc 120:10434–10440

    CAS  Google Scholar 

  50. Li H, Wurrey CJ, Thomas GJ (1992) J Am Chem Soc 114:7463–7469

    CAS  Google Scholar 

  51. Jarzecki AA, Spiro TG (2001) J Raman Spectrosc 32:599–605

    Article  CAS  Google Scholar 

  52. Brumlik MJ, Leroy G, Bruschi M, Voordouw G (1990) J Bacteriol 172:7289–7292

    CAS  PubMed  Google Scholar 

  53. Auchère F, Raleiras P, Benson L, Venyaminov SV, Tavares P, Moura JJG, Moura I, Rusnak F (2003) Inorg Chem 42:938–940

    Article  PubMed  Google Scholar 

  54. Bernstein HJ (2000) Trends Biochem Sci 25:453–455

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

This work was supported by grants from the National Institutes of Health (GM60329 to M.K.J. and GM40388 to D.M.K.) and a National Science Foundation Research Training Group Award to the Center for Metalloenzyme Studies (DBI9413236). We thank Radu Silaghi-Dumitrescu for helpful discussions.

Author information

Authors and Affiliations

  1. Department of Chemistry and Center for Metalloenzyme Studies, University of Georgia, Athens, GA , 30602, USA

    Michael D. Clay, Joseph P. Emerson, Eric D. Coulter, Donald M. Kurtz Jr. & Michael K. Johnson

Authors
  1. Michael D. Clay
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Joseph P. Emerson
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Eric D. Coulter
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Donald M. Kurtz Jr.
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Michael K. Johnson
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Michael K. Johnson.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Clay, M.D., Emerson, J.P., Coulter, E.D. et al. Spectroscopic characterization of the [Fe(His)4(Cys)] site in 2Fe-superoxide reductase from Desulfovibrio vulgaris . J Biol Inorg Chem 8, 671–682 (2003). https://doi.org/10.1007/s00775-003-0465-4

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00775-003-0465-4

Keywords

Search

Navigation