Development of 'Redox Arrays' for identifying novel glutathionylated proteins in the secretome

Mullen, Lisa, Seavill, Miles, Hammouz, Raneem, Bottazzi, Barbara, Chan, Philippe, Vaudry, David and Ghezzi, Pietro (2015) Development of 'Redox Arrays' for identifying novel glutathionylated proteins in the secretome. Scientific Reports, 5. p. 14630. ISSN 2045-2322

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Abstract

Proteomics techniques for analysing the redox status of individual proteins in complex mixtures tend to identify the same proteins due to their high abundance. We describe here an array-based technique to identify proteins undergoing glutathionylation and apply it to the secretome and the proteome of human monocytic cells. The method is based on incorporation of biotinylated glutathione (GSH) into proteins, which can then be identified following binding to a 1000-protein antibody array. We thus identify 38 secreted and 55 intracellular glutathionylated proteins, most of which are novel candidates for glutathionylation. Two of the proteins identified in these experiments, IL-1 sRII and Lyn, were then confirmed to be susceptible to glutathionylation. Comparison of the redox array with conventional proteomic methods confirmed that the redox array is much more sensitive, and can be performed using more than 100-fold less protein than is required for methods based on mass spectrometry. The identification of novel targets of glutathionylation, particularly in the secretome where the protein concentration is much lower, shows that redox arrays can overcome some of the limitations of established redox proteomics techniques.

Item Type: Article
Schools and Departments: Brighton and Sussex Medical School > Clinical and Experimental Medicine
Subjects: Q Science > QD Chemistry > QD0241 Organic chemistry > QD0415 Biochemistry
Q Science > QR Microbiology
Depositing User: Patricia Butler
Date Deposited: 27 Nov 2015 15:12
Last Modified: 05 Oct 2017 18:26
URI: http://sro.sussex.ac.uk/id/eprint/57977

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