Measurements for Sulfide-Mediated Inhibition of Myeloperoxidase Activity

  • Dorottya Garai
  • Zoltán Pálinkás
  • József Balla
  • Anthony J. Kettle
  • Péter NagyEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 2007)


Oxidative stress-alleviating and inflammation-mediatory functions of hydrogen sulfide were reported to be key features of its biological actions. However, the underlying molecular mechanisms of these biological observations are not fully understood. In conditions where sulfide was proposed to be protective against oxidative stress- or inflammation-induced tissue damage (e.g., reperfusion injury, atherosclerosis, vascular inflammation), the reactive oxidant-producing function of a key neutrophil enzyme, myeloperoxidase, was reported to be a protagonist on the detrimental side. We recently described favorable interactions between sulfide and myeloperoxidase and proposed that the potent inhibition of myeloperoxidase activities could contribute to sulfide’s beneficial functions in a number of cardiovascular pathologies. Our chapter is dedicated to aid future studies and drug development endeavors in this area by providing methodological guidance on how to assess the inhibitory potential of sulfide on myeloperoxidase enzymatic activities in isolated protein systems, in neutrophil homogenates, and in live neutrophil preparations.

Key words

Myeloperoxidase Hydrogen sulfide Peroxidase activity Halogenation activity Neutrophil function 





Dimethyl sulfoxide


Dulbecco's phosphate buffered saline


5,5-Dithio-bis-(2-nitrobenzoic acid)


Diethylenetriaminepentaacetic acid


Ethylenediaminetetraacetic acid


Hank’s Balanced Salt Solution


Horseradish peroxidase


Hexadecyltrimethylammonium bromide


Myeloperoxidase enzyme


Nicotinamide adenine dinucleotide phosphate (NADPH) oxidase 2 enzyme-complex


Phosphate-buffered saline


Phorbol 12-myristate 13-acetate


Relative fluorescence unit


Reactive oxygen species


Superoxide dismutase







This work was supported by The Hungarian National Science Foundation (OTKA; grant No.: K 109843, KH17_126766 and K18_129286 for P.N. and K 112333 for J.B.) and the National Institutes of Health (grant No.: R21AG055022-01 for P.N.). Financial supports from the Hungarian Government in a GINOP-2.3.2-15-2016-00043 project (for J.B. and P.N.) and from the European Union in a European Regional Development Fund are also acknowledged. The research group is supported by the Hungarian Academy of Sciences (11003). P.N. is a János Bolyai Research Scholar of the Hungarian Academy of Sciences. Dojindo Molecular Technologies Inc. is greatly acknowledged for their kind support of high-quality chemical supplies.


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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Dorottya Garai
    • 1
    • 2
  • Zoltán Pálinkás
    • 1
  • József Balla
    • 3
  • Anthony J. Kettle
    • 4
  • Péter Nagy
    • 1
    • 5
    Email author
  1. 1.Department of Molecular Immunology and ToxicologyNational Institute of OncologyBudapestHungary
  2. 2.Faculty of Medicine, Laki Kálmán Doctoral SchoolUniversity of DebrecenDebrecenHungary
  3. 3.HAS-UD Vascular Biology and Myocardial Pathophysiology Research GroupHungarian Academy of SciencesDebrecenHungary
  4. 4.Centre for Free Radical Research Department of Pathology and Biomedical ScienceUniversity of Otago ChristchurchChristchurchNew Zealand
  5. 5.Department of Medicine, Faculty of MedicineUniversity of DebrecenDebrecenHungary

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