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Measurements for Sulfide-Mediated Inhibition of Myeloperoxidase Activity

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Vascular Effects of Hydrogen Sulfide

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2007))

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Abstract

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.

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Abbreviations

DMF:

N,N-Dimethylformamide

DMSO:

Dimethyl sulfoxide

DPBS:

Dulbecco’s phosphate buffered saline

DTNB:

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

DTPA:

Diethylenetriaminepentaacetic acid

EDTA:

Ethylenediaminetetraacetic acid

HBSS:

Hank’s Balanced Salt Solution

HRP:

Horseradish peroxidase

HTAB:

Hexadecyltrimethylammonium bromide

MPO:

Myeloperoxidase enzyme

NOX2:

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

PBS:

Phosphate-buffered saline

PMA:

Phorbol 12-myristate 13-acetate

RFU:

Relative fluorescence unit

ROS:

Reactive oxygen species

SOD:

Superoxide dismutase

TMB:

3,3′,5,5′-Tetramethylbenzidine

UV-Vis:

Ultraviolet-visible

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Acknowledgments

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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Authors and Affiliations

  1. Department of Molecular Immunology and Toxicology, National Institute of Oncology, Budapest, Hungary

    Dorottya Garai, Zoltán Pálinkás & Péter Nagy

  2. Faculty of Medicine, Laki Kálmán Doctoral School, University of Debrecen, Debrecen, Hungary

    Dorottya Garai

  3. HAS-UD Vascular Biology and Myocardial Pathophysiology Research Group, Hungarian Academy of Sciences, Debrecen, Hungary

    József Balla

  4. Centre for Free Radical Research Department of Pathology and Biomedical Science, University of Otago Christchurch, Christchurch, New Zealand

    Anthony J. Kettle

  5. Department of Medicine, Faculty of Medicine, University of Debrecen, Debrecen, Hungary

    Péter Nagy

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  1. Dorottya Garai
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  2. József Balla
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  3. Anthony J. Kettle
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  4. Péter Nagy
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Correspondence to Péter Nagy .

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  1. Department of Pathophysiology, Medical University, Lublin, Poland

    Jerzy Bełtowski

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Garai, D., Pálinkás, Z., Balla, J., Kettle, A.J., Nagy, P. (2019). Measurements for Sulfide-Mediated Inhibition of Myeloperoxidase Activity. In: Bełtowski, J. (eds) Vascular Effects of Hydrogen Sulfide. Methods in Molecular Biology, vol 2007. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9528-8_14

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  • DOI: https://doi.org/10.1007/978-1-4939-9528-8_14

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-9527-1

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