Annulé | Dr. Claire Eyers - Phosphorylation-mediated signalling: a new era?

Claire Eyers

University of Liverpool, Liverpool, UK

Phosphorylation-mediated signalling: a new era?

Protein phosphorylation is a ubiquitous post-translational modification (PTM) that regulates all aspects of life. Using our novel UPAX methodology for unbiased phosphopeptide enrichment and analysis by LC/MSMS, we demonstrate that human proteins are phosphorylated extensively on residues in addition to serine, threonine and tyrosine. Of particular interest is phosphorylation of cysteine, and the implications of this novel protein modification as a regulatory signalling PTM, which we show of relevance to PINK1 and HIF signalling. Cys is also known to be regulated through oxidation and we will explore some of our findings in the area of Cys oxidation linked to the regulation of protein kinase activity.
Selected references

• Considerations for defining +80 Da mass shifts in mass spectrometry-based proteomics: phosphorylation and beyond. Daly LA, Clarke CJ, Po A, Oswald SO, Eyers CE. Chem Commun (Camb) 2023 Sep 26;59(77):11484-11499
• Mutation of the RelA(p65) Thr505 phosphosite disrupts the DNA replication stress response leading to CHK1 inhibitor resistance. Hunter JE, Campbell AE, Butterworth JA, Sellier H, Hannaway NL, Luli S, Floudas A, Kenneth NS, Moore AJ, Brownridge PJ, Thomas HD, Coxhead J, Taylor L, Leary P, Hasoon MSR, Knight AM, Garrett MD, Collins I, Eyers CE, Perkins ND. Biochem J 2022 Oct 14;479(19):2087-2113
• Oxygen-dependent changes in binding partners and post-translational modifications regulate the abundance and activity of HIF-1α/2α. Daly LA, Brownridge PJ, Batie M, Rocha S, Sée V, Eyers CE. Sci Signal 2021 Jul 20;14(692):eabf6685
• Temporal modulation of the NF-κB RelA network in response to different types of DNA damage. Campbell AE, Ferraz Franco C, Su LI, Corbin EK, Perkins S, Kalyuzhnyy A, Jones AR, Brownridge PJ, Perkins ND, Eyers CE. Biochem J 2021 Feb 12;478(3):533-551
• Use of the Polo-like kinase 4 (PLK4) inhibitor centrinone to investigate intracellular signalling networks using SILAC-based phosphoproteomics. Byrne DP, Clarke CJ, Brownridge PJ, Kalyuzhnyy A, Perkins S, Campbell A, Mason D, Jones AR, Eyers PA, Eyers CE. Biochem J 2020 Jul 17;477(13):2451-2475
• Strong anion exchange-mediated phosphoproteomics reveals extensive human non-canonical phosphorylation. Hardman G, Perkins S, Brownridge PJ, Clarke CJ, Byrne DP, Campbell AE, Kalyuzhnyy A, Myall A, Eyers PA, Jones AR, Eyers CE. EMBO J 2019 Oct 4;38(21):e100847