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Characterization of the copper(II) binding site in the pink copper binding protein CusF by electron paramagnetic resonance spectroscopy

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Abstract

Electron paramagnetic resonance (EPR) spectroscopy has been used to structurally characterize the copper-binding site in CusF protein from Escherichia coli. The EPR spectra indicate a single type II copper center with parameters typical for nitrogen and oxygen ligands (A~200 G, g~2.186, g~2.051). The pulsed EPR data show that one of the ligands to Cu2+ is an imidazole ring of a histidine residue. The remote amino nitrogen of this imidazole ring is readily observed by electron spin-echo envelope modulation spectroscopy, while the imino nitrogen that is directly coordinated to the Cu2+ ion is observed by pulsed electron–nuclear double resonance (ENDOR). In addition, the ENDOR spectra reveal the presence of one more nitrogen ligand that was assigned to be a deprotonated peptide nitrogen. Apart from the two nitrogen ligands, it has been established that there are two nearby hydroxyl protons, although whether these belong to a single equatorial water ligand or two equatorial hydroxide ligands is not known.

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Notes

  1. The Euler angles as used in this work specify three consecutive rotations in the following order: (1) by φ around axis 3 (or z); (2) by θ around newly obtained axis 2 (or y); (3) by ψ around newly obtained axis 3 (or z).

References

  1. Franke S, Grass G, Nies DH (2001) Microbiology 147:965–972

    CAS  PubMed  Google Scholar 

  2. Franke S, Grass G, Rensing C, Nies DH (2003) J Bacteriol 185:3804–3812

    Article  PubMed  Google Scholar 

  3. Rensing C, Grass G (2003) FEMS Microbiol Rev 27:197–213

    Article  CAS  PubMed  Google Scholar 

  4. Gupta A, Phung LT, Taylor DE, Silver S (2001) Microbiology 147:3393–3402

    CAS  PubMed  Google Scholar 

  5. Astashkin AV, Raitsimring AM, Kennedy AR, Shokhireva TK, Walker FA (2002) J Phys Chem A 106:74–82

    Google Scholar 

  6. Peisach J, Blumberg WE (1974) Arch Biochem Biophys 165:691–708

    CAS  PubMed  Google Scholar 

  7. Dikanov SA, Tsvetkov YD (1992) Electron spin echo envelope modulation (ESEEM) spectroscopy. CRC Press, Boca Raton

    Google Scholar 

  8. Edmonds DT (1977) Phys Rep 29:234–290

    Google Scholar 

  9. Mims WB (1965) Proc R Soc Lond Ser-A 283:452

    Google Scholar 

  10. Davies ER (1974) Phys Lett A 47:1–2

    Google Scholar 

  11. Vancamp HL, Sands RH, Fee JA (1981) J Chem Phys 75:2098–2107

    Google Scholar 

  12. Polt R, Kelly BD, Dangel BD, Tadikonda UB, Ross RE, Raitsimring AM, Astashkin AV (2003) Inorg Chem 42:566–574

    Google Scholar 

  13. Roberts JE, Cline JF, Lum V, Freeman H, Gray HB, Peisach J, Reinhammar B, Hoffman BM (1984) J Am Chem Soc 106:5324–5330

    Google Scholar 

  14. Iwaizumi M, Kudo T, Kita S (1986) Inorg Chem 25:1546–1550

    Google Scholar 

  15. Raitsimring AM, Pacheco A, Enemark JH (1998) J Am Chem Soc 120:11263–11278

    Google Scholar 

  16. Raitsimring AM, Astashkin AV, Feng CJ, Enemark JH, Nelson KJ, Rajagopalan KV (2003) J Biol Inorg Chem 8:95–104

    Google Scholar 

  17. Semin GK, Babushkina TA, Iakobson GG (1975) Nuclear quadrupole resonance in chemistry. Wiley, New York

    Google Scholar 

  18. Atherton NM, Horsewill AJ (1979) Mol Phys 37:1349–1361

    Google Scholar 

  19. Sigel H, Martin RB (1982) Chem Rev 82:385–426

    Google Scholar 

  20. Zhidomirov GI, Schastnev PV, Chuvylkin ND (1978) Quantum-chemical calculations of magnetic resonance parameters. Nauka, Novosibirsk

    Google Scholar 

  21. Ashby CIH, Cheng CP, Duesler EN, Brown TL (1978) J Am Chem Soc 100:6063–6067

    Google Scholar 

  22. Freeman HC, Taylor MR (1965) Acta Crystallogr 18:939

    Google Scholar 

  23. Diaddario LL, Robinson WR, Margerum DW (1983) Inorg Chem 22:1021–1025

    Google Scholar 

  24. Getz D, Silver BL (1974) J Chem Phys 61:630–637

    Google Scholar 

  25. Harford C, Sarkar B (1997) Acc Chem Res 30:123–130

    Google Scholar 

  26. Prenesti E, Daniele PG, Prencipe M, Ostacoli G (1999) Polyhedron 18:3233–3241

    Google Scholar 

  27. Castagnetto JM, Hennessy SW, Roberts VA, Getzoff ED, Tainer JA, Pique ME (2002) Nucleic Acids Res 30:379–382

    Google Scholar 

  28. Zaitseva I, Zaitsev V, Card G, Moshkov K, Bax B, Ralph A, Lindley P (1996) J Biol Inorg Chem 1:15–23

    Google Scholar 

  29. Utschig LM, Astashkin AV, Raitsimring AM, Thurnauer MC, Poluektov OG (2004) J Phys Chem B 108:11150–11156

    Google Scholar 

  30. Colaneri MJ, Peisach J (1995) J Am Chem Soc 117:6308–6315

    Google Scholar 

Download references

Acknowledgements

We thank Joshua Kittleson for assistance with sample preparation. The authors acknowledge the NSF funding (DBI-9604939) for construction and modification of the pulsed EPR spectrometer used in this work.

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Correspondence to Megan M. McEvoy.

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Astashkin, A.V., Raitsimring, A.M., Walker, F.A. et al. Characterization of the copper(II) binding site in the pink copper binding protein CusF by electron paramagnetic resonance spectroscopy. J Biol Inorg Chem 10, 221–230 (2005). https://doi.org/10.1007/s00775-005-0631-y

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  • DOI: https://doi.org/10.1007/s00775-005-0631-y

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