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Biophysics

, Volume 53, Issue 4, pp 268–272 | Cite as

Influence of ceruloplasmin and lactoferrin on the chlorination activity of leukocyte myeloperoxidase assayed by chemiluminescence

  • O. M. PanasenkoEmail author
  • A. V. Chekanov
  • I. I. Vlasova
  • A. V. Sokolov
  • K. V. Ageeva
  • M. O. Pulina
  • O. S. Cherkalina
  • V. B. Vasil’ev
Molecular Biophysics

Abstract

We demonstrate that addition of H2O2 to a mixture of myeloperoxidase (MPO), chloride and luminol immediately evokes a short intense flash of chemiluminescence (CL). This flash is diminished in the absence of MPO or chloride, and in the complete system it is suppressed by an MPO inhibitor azide, hypochlorite scavengers taurine or methionine, or an MPO peroxidase-cycle substrate guaiacol. Hence, this CL is mostly due to the MPO halogenation function; a measure of this activity is provided by the integral CL. With three independent methods (CL, taurine chlorination, and peroxidase assay) it is shown that MPO activity is suppressed by ceruloplasmin (Cp). Lactoferrin has no effect either on MPO or on the MPO-Cp complex. It is also shown that peroxidase inhibition by Cp is the stronger the larger is the MPO substrate, which suggests steric hindrances to substrate binding in the MPO-Cp complex. Importantly, the conventional chlorination and peroxidase assays detect MPO inhibition by Cp only at a large excess of the latter, whereas the CL assay reveals it at stoichiometric ratios characteristic of the naturally occurring protein complexes.

Key words

myeloperoxidase ceruloplasmin hypochlorite chemiluminescence protein-protein interactions 

Abbreviations

ABTS

sodium 2,2′-azinobis(3-ethylbenzotriazoline-6-sulfonate

CL

chemiluminescence

Cp

ceruloplasmin

Lf

lactoferrin

MPO

myeloperoxidase

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Copyright information

© Pleiades Publishing, Ltd. 2008

Authors and Affiliations

  • O. M. Panasenko
    • 1
    Email author
  • A. V. Chekanov
    • 1
  • I. I. Vlasova
    • 1
  • A. V. Sokolov
    • 2
  • K. V. Ageeva
    • 2
  • M. O. Pulina
    • 2
  • O. S. Cherkalina
    • 2
  • V. B. Vasil’ev
    • 2
  1. 1.Research Institute of Physico-Chemical MedicineMinistry of Health of the Russian FederationMoscowRussia
  2. 2.Institute of Experimental MedicineRussian Academy of Medical SciencesSt. PetersburgRussia

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