Skip to main content
SpringerLink
Log in

Fluoride and azide binding to ferric human hemoglobin:haptoglobin complexes highlights the ligand-dependent inequivalence of the α and β hemoglobin chains

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

Abstract

Haptoglobin (Hp) binds human hemoglobin (Hb), contributing to prevent extra-erythrocytic Hb-induced damage. Hp forms preferentially complexes with αβ dimers, displaying heme-based reactivity. Here, kinetics and thermodynamics of fluoride and azide binding to ferric human Hb (Hb(III)) complexed with the human Hp phenotypes 1-1 and 2-2 (Hp1-1:Hb(III) and Hp2-2:Hb(III), respectively) are reported (pH 7.0 and 20.0 °C). Fluoride binds to Hp1-1:Hb(III) and Hp2-2:Hb(III) with a one-step kinetic and equilibrium behavior. In contrast, kinetics of azide binding to and dissociation from Hp1-1:Hb(III)(–N3−) and Hp2-2:Hb(III)(–N3−) follow a two-step process. However, azide binding to Hp1-1:Hb(III) and Hp2-2:Hb(III) is characterized by a simple equilibrium, reflecting the compensation of kinetic parameters. The fast and the slow step of azide binding to Hp1-1:Hb(III) and Hp2-2:Hb(III) should reflect azide binding to the ferric β and α chains, respectively, as also proposed for the similar behavior observed in Hb(III). Present results highlight the ligand-dependent kinetic inequivalence of Hb subunits in the ferric form, reflecting structural differences between the two subunits in the interaction with some ferric ligands.

Graphical abstract

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

Scheme 1
Scheme 2
Fig. 1
Fig. 2

Similar content being viewed by others

Abbreviations

CCP domain:

Complement control protein domain

Hb:

Human hemoglobin

Hb(III):

Ferric Hb

Hp:

Human haptoglobin

Hp1-1:

Phenotype 1-1 of Hp

Hp2-2:

Phenotype 2-2 of Hp

Hp1-1:Hb(III):

Ferric Hp1-1:Hb complex

Hp2-2:Hb(III):

Ferric Hp 2-2:Hb complex

SP-like domain:

Serine protease-like domain

References

  1. Muller-Eberhard U, Javid J, Liem HH, Hanstein A, Hanna M (1968) Blood 32:811–815

    PubMed  CAS  Google Scholar 

  2. Bunn HF, Forget BG (1986) Hemoglobin: molecular, genetic and clinical aspects. Saunders, Philadelphia

    Google Scholar 

  3. Ascenzi P, Bocedi A, Visca P, Altruda F, Tolosano E, Beringhelli T, Fasano M (2005) IUBMB Life 57:749–759

    Article  PubMed  CAS  Google Scholar 

  4. Alayash AI, Andersen CB, Moestrup SK, Bülow L (2013) Trends Biotechnol 31:2–3

    Article  PubMed  CAS  Google Scholar 

  5. Alayash AI (2004) Nat Rev Drug Discov 3:152–159

    Article  PubMed  CAS  Google Scholar 

  6. Schaer DJ, Vinchi F, Ingoglia G, Tolosano E, Buehler PW (2014) Front Physiol 5:415

    Article  PubMed  PubMed Central  Google Scholar 

  7. MacKellar M, Vigerust DJ (2016) Clin Diabetes 34:148–157

    Article  PubMed  PubMed Central  Google Scholar 

  8. Kristiansen M, Graversen JH, Jacobsen C, Sonne O, Hoffman HJ, Law SK, Moestrup SK (2001) Nature 409:198–201

    Article  PubMed  CAS  Google Scholar 

  9. Buehler PW, Abraham B, Vallelian F, Linnemayr C, Pereira CP, Cipollo JF, Jia Y, Mikolajczyk M, Boretti FS, Schoedon G, Alayash AI, Schaer DJ (2009) Blood 113:2578–2586

    Article  PubMed  CAS  Google Scholar 

  10. Kaempfer T, Duerst E, Gehrig P, Roschitzki B, Rutishauser D, Grossmann J, Schoedon G, Vallelian F, Schaer DJ (2011) J Proteome Res 10:2397–2408

    Article  PubMed  CAS  Google Scholar 

  11. Andersen CBF, Stødkilde K, Sæderup KL, Kuhlee A, Raunser S, Graversen JH, Moestrup SK (2017) Antioxid Redox Signal 26:814–831

    Article  PubMed  CAS  Google Scholar 

  12. Polticelli F, Bocedi A, Minervini G, Ascenzi P (2008) FEBS J 275:5648–5656

    Article  PubMed  CAS  Google Scholar 

  13. Andersen CB, Torvund-Jensen M, Nielsen MJ, de Oliveira CL, Hersleth HP, Andersen NH, Pedersen JS, Andersen GR, Moestrup SK (2012) Nature 489:456–459

    Article  PubMed  CAS  Google Scholar 

  14. Stødkilde K, Torvund-Jensen M, Moestrup SK, Andersen CB (2014) Nat Commun 5:5487

    Article  PubMed  CAS  Google Scholar 

  15. Kurosky A, Barnett DR, Lee TH, Touchstone B, Hay RE, Arnott MS, Bowman BH, Fitch WM (1980) Proc Natl Acad Sci USA 77:3388–3392

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  16. Wejman JC, Hovsepian D, Wall JS, Hainfeld JF, Greer J (1984) J Mol Biol 174:319–341

    Article  PubMed  CAS  Google Scholar 

  17. Wejman JC, Hovsepian D, Wall JS, Hainfeld JF, Greer J (1984) J Mol Biol 174:343–368

    Article  PubMed  CAS  Google Scholar 

  18. Nagel RL, Gibson QH (1971) J Biol Chem 246:69–73

    PubMed  CAS  Google Scholar 

  19. Nagel RL, Wittenberg JB, Ranney HM (1965) Biochim Biophys Acta 100:286–289

    Article  PubMed  CAS  Google Scholar 

  20. Nagel RL, Gibson QH (1966) J Mol Biol 22:249–255

    Article  PubMed  CAS  Google Scholar 

  21. Brunori M, Alfsen A, Saggese U, Antonini E, Wyman J (1968) J Biol Chem 243:2950–2954

    PubMed  CAS  Google Scholar 

  22. Gibson QH, Parkhurst LJ, Geraci G (1969) J Biol Chem 244:4668–4676

    PubMed  CAS  Google Scholar 

  23. Alfsen A, Chiancone E, Antonini E, Waks M, Wyman J (1970) Biochim Biophys Acta 207:395–403

    Article  PubMed  CAS  Google Scholar 

  24. Antonini E, Brunori M (1971) Hemoglobin and myoglobin in their reactions with ligands. North Holland Publishing Co, Amsterdam

    Google Scholar 

  25. Chiancone E, Antonini E, Brunori M, Alfsen A, Lavialle F (1973) Biochem J 133:205–207

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  26. Perutz MF (1979) Annu Rev Biochem 48:327–386

    Article  PubMed  CAS  Google Scholar 

  27. Azarov I, He X, Jeffers A, Basu S, Ucer B, Hantgan RR, Levy A, Kim-Shapiro DB (2008) Nitric Oxide 18:296–302

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  28. Ascenzi P, Tundo GR, Coletta M (2018) J Inorg Biochem 187:116–122

    Article  PubMed  CAS  Google Scholar 

  29. Coletta M, Angeletti M, De Sanctis G, Cerroni L, Giardina B, Amiconi G, Ascenzi P (1996) Eur J Biochem 235:49–53

    Article  PubMed  CAS  Google Scholar 

  30. Ackers GK, Doyle ML, Myers D, Daugherty MA (1992) Science 255:54–63

    Article  PubMed  CAS  Google Scholar 

  31. White SL (1975) J Biol Chem 250:1263–1268

    PubMed  CAS  Google Scholar 

  32. Ascenzi P, De Simone G, Polticelli F, Gioia M, Coletta M (2018) J Biol Inorg Chem 23:437–445

    Article  PubMed  CAS  Google Scholar 

  33. Ascenzi P, Coletta M (2018) J Phys Chem B 122:11100–11107

    Article  CAS  PubMed  Google Scholar 

  34. Herold S, Shivashankar K (2003) Biochemistry 42:14036–14046

    Article  PubMed  CAS  Google Scholar 

  35. Klapper MH, Uchida H (1971) J Biol Chem 246:6849–6854

    PubMed  CAS  Google Scholar 

  36. Deatherage JF, Loe RS, Moffat K (1976) J Mol Biol 104:723–728

    Article  PubMed  CAS  Google Scholar 

  37. Deatherage JF, Obendorf SK, Moffat K (1979) J Mol Biol 134:419–429

    Article  PubMed  CAS  Google Scholar 

  38. Banerjee S, Jia Y, Siburt CJ, Abraham B, Wood F, Bonaventura C, Henkens R, Crumbliss AL, Alayash AI (2012) Free Radic Biol Med 53:1317–1326

    Article  PubMed  CAS  Google Scholar 

  39. Anusiem AC, Beetlestone JG, Irvine DH (1968) J Chem Soc A 960–969

  40. Bailey JE, Beetlestone JG, Irvine DH (1968) J Chem Soc A 2778–2783

Download references

Acknowledgements

The Grant of Dipartimenti di Eccellenza, MIUR (Legge 232/2016, Articolo 1, Comma 314-337) is gratefully acknowledged.

Author information

Authors and Affiliations

  1. Interdepartmental Laboratory for Electron Microscopy, Roma Tre University, Via della Vasca Navale 79, 00146, Roma, Italy

    Paolo Ascenzi

  2. Department of Sciences, Roma Tre University, Viale Marconi 446, 00146, Roma, Italy

    Alessandra di Masi & Giovanna De Simone

  3. Department of Clinical Sciences and Translational Medicine, University of Roma “Tor Vergata”, Via Montpellier 1, 00133, Roma,, Italy

    Magda Gioia & Massimo Coletta

  4. Interuniversity Consortium for the Research on the Chemistry of Metals in Biological Systems, Via Celso Ulpiani 27, 70126, Bari, Italy

    Magda Gioia & Massimo Coletta

Authors
  1. Paolo Ascenzi
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Alessandra di Masi
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Giovanna De Simone
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Magda Gioia
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Massimo Coletta
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Paolo Ascenzi.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Ascenzi, P., di Masi, A., De Simone, G. et al. Fluoride and azide binding to ferric human hemoglobin:haptoglobin complexes highlights the ligand-dependent inequivalence of the α and β hemoglobin chains. J Biol Inorg Chem 24, 247–255 (2019). https://doi.org/10.1007/s00775-019-01642-9

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00775-019-01642-9

Keywords

Search

Navigation