Skip to main content
SpringerLink
Log in

Co(II) is not oxidized during turnover in the copper amine oxidase from Hansenula polymorpha

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

Abstract

Co(II) substitution into the copper amine oxidases (CAOs) has been an effective tool for evaluating the mechanism of oxygen reduction in these enzymes. However, formation of hydrogen peroxide during turnover raises questions about the relevant oxidation state of the cobalt in these enzymes and, therefore, the interpretation of the activity of the metal-substituted enzyme with respect to its mechanism of action. In this study, Co(II) was incorporated into the CAO from Hansenula polymorpha (HPAO). The effect of hydrogen peroxide on the catalytic activity of cobalt-substituted HPAO was evaluated. Hydrogen peroxide, either generated during turnover or added exogenously, caused a decrease in the activity of the enzyme but did not oxidize Co(II) to Co(III). These results are in strong contrast with results from the CAO from Arthrobacter globiformis (AGAO), where hydrogen peroxide causes an increase in the activity of the enzyme as the Co(II) is oxidized to Co(III). The results of this study with HPAO support previous reports that have shown that this enzyme acts by transferring an electron directly from the reduced TPQ cofactor to dioxygen rather than passing the electron through the bound metal ion. Furthermore, these results provide additional evidence to support the idea that different CAOs use different mechanisms for catalysis.

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.

Scheme 1
Scheme 2
Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

Abbreviations

AGAO:

Arthrobacter globiformis amine oxidase

CAO:

Copper amine oxidase

CoHPAO:

Cobalt-substituted HPAO

CuDepHPAO:

Copper-depleted HPAO

EPR:

Electron paramagnetic resonance

HPAO:

Hansenula polymorpha amine oxidase

ICP-AES:

Inductively coupled plasma-atomic emission spectroscopy

MOPS:

3-(N-Morpholino)-propanesulfonic acid

PHZ:

Phenylhydrazine

PSAO:

Pea seedling amine oxidase

TPQ:

2,4,5-Trihydroxyphenylalanyl quinone

References

  1. McIntire WS, Hartmann C (1993) Copper-containing amine oxidases. In: Davidson VL (ed) Principles and applications of quinoproteins. Marcel Dekker Inc, New York, pp 97–171

    Google Scholar 

  2. Klinman JP (2003) The multi-functional topa-quinone copper amine oxidases. Biochim Biophys Acta 1647:131–137

    Article  CAS  PubMed  Google Scholar 

  3. Mure M, Mills SA, Klinman JP (2002) Catalytic mechanism of the topa quinone containing copper amine oxidases. Biochemistry 41:9269–9278

    Article  CAS  PubMed  Google Scholar 

  4. Mure M (2004) Tyrosine-derived quinone cofactors. Acc Chem Res 37:131–139. https://doi.org/10.1021/ar9703342

    Article  CAS  PubMed  Google Scholar 

  5. Mills SA, Klinman JP (2000) Evidence against reduction of Cu2+ to Cu+ during dioxygen activation in a copper amine oxidase from yeast. J Am Chem Soc 122:9897–9904

    Article  CAS  Google Scholar 

  6. Su Q, Klinman JP (1998) Probing the mechanism of proton coupled electron transfer to dioxygen: the oxidative half-reaction of bovine serum amine oxidase. Biochemistry 37:12513–12525. https://doi.org/10.1021/bi981103l

    Article  CAS  PubMed  Google Scholar 

  7. Dooley DM, McGuirl MA, Brown DE et al (1991) A Cu(I)-semiquinone state in substrate-reduced amine oxidases. Nature 349:262–264. https://doi.org/10.1038/349262a0

    Article  CAS  PubMed  Google Scholar 

  8. Mukherjee A, Smirnov VV, Lanci MP et al (2008) Inner-sphere mechanism for molecular oxygen reduction catalyzed by copper amine oxidases. J Am Chem Soc 130:9459–9473. https://doi.org/10.1021/ja801378f

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Mills S, Brown D, Dang K et al (2012) Cobalt substitution supports an inner-sphere electron transfer mechanism for oxygen reduction in pea seedling amine oxidase. J Biol Inorg Chem 17:507–515. https://doi.org/10.1007/s00775-011-0872-x

    Article  CAS  PubMed  Google Scholar 

  10. Turowski PN, McGuirl MA, Dooley DM (1993) Intramolecular electron transfer rate between active-site copper and topa quinone in pea seedling amine oxidase. J Biol Chem 268:17680–17682

    CAS  PubMed  Google Scholar 

  11. Dooley DM, Brown DE (1996) Intramolecular electron transfer in the oxidation of amines by methylamine oxidase from Arthrobacter P1. J Biol Inorg Chem 1:205–209. https://doi.org/10.1007/s007750050044

    Article  CAS  Google Scholar 

  12. Shepard EM, Okonski KM, Dooley DM (2008) Kinetics and spectroscopic evidence that the Cu(I)-semiquinone intermediate reduces molecular oxygen in the oxidative half-reaction of Arthrobacter globiformis amine oxidase. Biochemistry 47:13907–13920. https://doi.org/10.1021/bi8011516

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Shepard EM, Dooley DM (2006) Intramolecular electron transfer rate between active-site copper and TPQ in Arthrobacter globiformis amine oxidase. J Biol Inorg Chem 11:1039–1048. https://doi.org/10.1007/s00775-006-0153-2

    Article  CAS  PubMed  Google Scholar 

  14. Liu Y, Mukherjee A, Nahumi N et al (2013) Experimental and computational evidence of metal-O2 activation and rate-limiting proton-coupled electron transfer in a copper amine oxidase. J Phys Chem B 117:218–229. https://doi.org/10.1021/jp3121484

    Article  CAS  PubMed  Google Scholar 

  15. Mills SA, Goto Y, Su QJ et al (2002) Mechanistic comparison of the cobalt-substituted and wild-type copper amine oxidase from Hansenula polymorpha. Biochemistry 41:10577–10584. https://doi.org/10.1021/bi0200864

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

SAM acknowledges the other Xavier students who work in the lab. SAM and KEG acknowledge the Xavier University Office of Undergraduate Research for support.

Author information

Authors and Affiliations

  1. Department of Chemistry, Xavier University, 3800 Victory Parkway, Cincinnati, OH, 45207, USA

    Stephen A. Mills & Kiera E. Gazica

  2. Department of Chemistry and Biochemistry, Miami University, 651 E. High St, Oxford, OH, 45056, USA

    David L. Tierney

Authors
  1. Stephen A. Mills
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Kiera E. Gazica
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. David L. Tierney
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Stephen A. Mills.

Additional information

This work is dedicated to Prof. Justine P. Roth, who was a brilliant scientist and great sounding board for ideas.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Mills, S.A., Gazica, K.E. & Tierney, D.L. Co(II) is not oxidized during turnover in the copper amine oxidase from Hansenula polymorpha. J Biol Inorg Chem 24, 31–37 (2019). https://doi.org/10.1007/s00775-018-1624-y

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00775-018-1624-y

Keywords

Search

Navigation