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Detection of Nitric Oxide by Electron Paramagnetic Resonance Spectroscopy: Spin-Trapping with Iron-Dithiocarbamates

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Plant Nitric Oxide

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1424))

Abstract

Electron paramagnetic resonance (EPR) spectroscopy is the ideal methodology to identify radicals (detection and characterization of molecular structure) and to study their kinetics, in both simple and complex biological systems. The very low concentration and short life-time of NO and of many other radicals do not favor its direct detection and spin-traps are needed to produce a new and persistent radical that can be subsequently detected by EPR spectroscopy.

In this chapter, we present the basic concepts of EPR spectroscopy and of some spin-trapping methodologies to study NO. The “strengths and weaknesses” of iron-dithiocarbamates utilization, the NO traps of choice for the authors, are thoroughly discussed and a detailed description of the method to quantify the NO formation by molybdoenzymes is provided.

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Abbreviations

AOR:

Aldehyde oxidoreductase

CPTIO:

2-(4-Carboxyphenyl)-4,4,5,5-tetramethyl-imidazoline-1-oxyl-3-oxide

DETC:

Diethyldithiocarbamate

DMPO:

5,5-Dimethyl-1-pyrroline N-oxide

EPR:

Electron paramagnetic resonance

Fe-(TC)2 :

Iron-bis-dithiocarbamates

Fe2+-(TC)2 :

Ferrous-bis-dithiocarbamate

Fe3+-(TC)2 :

Ferric-bis-dithiocarbamate

Hb:

Hemoglobin

MGD:

N-methyl-d-glucamine-dithiocarbamate

MNIC:

Mononitrosyl-iron complex

MNP:

2-Methyl-2-nitrosopropane

NO:

Nitric oxide radical (•NO)

ST:

Spin-trap molecule

TEMPO:

2,2,6,6-Tetramethylpiperidinyl-N-oxyl

XO:

Xanthine oxidase

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Acknowledgments

This work was supported by the Unidade de Ciências Biomoleculares Aplicadas-UCIBIO which is financed by national funds from FCT/MEC (UID/Multi/04378/2013) and co-financed by the ERDF under the PT2020 Partnership Agreement (POCI-01-0145-FEDER-007728.

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Authors and Affiliations

  1. UCIBIO, REQUIMTE, Departamento Química, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, Campus de Caparica, 2829-516, Caparica, Portugal

    Luisa B. Maia & José J. G. Moura

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  1. Luisa B. Maia
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  2. José J. G. Moura
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Correspondence to Luisa B. Maia .

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  1. National Institute of Plant Genome Resea, New Delhi, India

    Kapuganti Jagadis Gupta

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Maia, L.B., Moura, J.J.G. (2016). Detection of Nitric Oxide by Electron Paramagnetic Resonance Spectroscopy: Spin-Trapping with Iron-Dithiocarbamates. In: Gupta, K. (eds) Plant Nitric Oxide. Methods in Molecular Biology, vol 1424. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3600-7_8

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  • DOI: https://doi.org/10.1007/978-1-4939-3600-7_8

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