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Investigating ROS, RNS, and H2S-sensitive signaling proteins

Williams, Eleanor; Whiteman, Matthew; Wood, Mark E.; Wilson, Ian D.; Ladomery, Michael R.; Allainguillaume, Joel; Teklic, Tihana; Lisjak, Miro; Hancock, John T.


Eleanor Williams

Matthew Whiteman

Mark E. Wood

Ian D. Wilson

Tihana Teklic

Miro Lisjak

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John Hancock
Professor in Cell Signalling



© Springer Science+Business Media, LLC, part of Springer Nature 2019. The modification of proteins is a key way to alter their activity and function. Often thiols, cysteine residues, on proteins are attractive targets for such modification. Assuming that the thiol group is accessible then reactions may take place with a range of chemicals found in cells. These may include reactive oxygen species (ROS), such as hydrogen peroxide (H2O2), reactive nitrogen species such as nitric oxide (NO), hydrogen sulfide (H2S), or glutathione. Such modifications often are instrumental to important cellular signaling processes, which ultimately result in modification of physiology of the organism. Therefore, there is a need to be able to identify such modifications. There are a variety of techniques to find proteins which may be altered in this way but here the focus is on two approaches: Firstly, the use of fluorescent thiol derivatives and the subsequent use of mass spectrometry to identify the thiols involved; secondly the confirmation of such changes using biochemical assays and genetic mutants. The discussion will be based on the use of two model organisms: Firstly the plant Arabidopsis thaliana (both as cell cultures and whole plants) and secondly the nematode worm Caenorhabditis elegans. However, these tools, as described, may be used in a much wider range of biological systems, including human and human tissue cultures.


Williams, E., Whiteman, M., Wood, M. E., Wilson, I. D., Ladomery, M. R., Allainguillaume, J., …Hancock, J. T. (2019). Investigating ROS, RNS, and H2S-sensitive signaling proteins. In J. T. Hancock, & M. E. Conway (Eds.), Redox-Mediated Signal Transduction (27-42). Humana, New York, NY.

Online Publication Date May 31, 2019
Publication Date 2019
Journal Methods in Molecular Biology
Peer Reviewed Not Peer Reviewed
Volume 1990
Pages 27-42
Series Title Methods in Molecular Biology
Series Number 1990
Book Title Redox-Mediated Signal Transduction
ISBN 9781493994618
Keywords glutathione, glyceraldehyde 3-phosphate dehydrogenase, histidine kinase, hydrogen peroxide, hydrogen sulfide, 5′-iodoacetamide fluorescein, nitric oxide, reactive nitrogen species, reactive oxygen species, stomatal guard cells, thiol labeling
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