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Exercise decreases PP2A-specific reversible thiol oxidation in human erythrocytes: Implications for redox biomarkers

Muggeridge, David J.; Crabtree, Daniel R.; Tuncay, Ahmet; Megson, Ian L.; Davison, Gareth; Cobley, James N.


Daniel R. Crabtree

Ahmet Tuncay

Ian L. Megson

Gareth Davison

James N. Cobley


New readily accessible systemic redox biomarkers are needed to understand the biological roles reactive oxygen species (ROS) play in humans because overtly flawed, technically fraught, and unspecific assays severely hamper translational progress. The antibody-linked oxi-state assay (ALISA) makes it possible to develop valid ROS-sensitive target-specific protein thiol redox state biomarkers in a readily accessible microplate format. Here, we used a maximal exercise bout to disrupt redox homeostasis in a physiologically meaningful way to determine whether the catalytic core of the serine/threonine protein phosphatase PP2A is a candidate systemic redox biomarker in human erythrocytes. We reasoned that: constitutive oxidative stress (e.g., haemoglobin autoxidation) would sensitise erythrocytes to disrupted ion homeostasis as manifested by increased oxidation of the ion regulatory phosphatase PP2A. Unexpectedly, an acute bout of maximal exercise lasting ˜16 min decreased PP2A-specific reversible thiol oxidation (redox ratio, rest: 0.46; exercise: 0.33) without changing PP2A content (rest: 193 pg/ml; exercise: 191 pg/ml). The need for only 3–4 μl of sample to perform ALISA means PP2A-specific reversible thiol oxidation is a capillary—fingertip blood—compatible candidate redox biomarker. Consistent with biologically meaningful redox regulation, thiol reductant-inducible PP2A activity was significantly greater (+10%) at rest compared to exercise. We establish a route to developing new readily measurable protein thiol redox biomarkers for understanding the biological roles ROS play in humans.

Journal Article Type Article
Acceptance Date Feb 18, 2022
Online Publication Date Feb 22, 2022
Publication Date 2022-03
Deposit Date Feb 23, 2022
Publicly Available Date Feb 23, 2023
Journal Free Radical Biology and Medicine
Print ISSN 0891-5849
Publisher Elsevier
Peer Reviewed Peer Reviewed
Volume 182
Pages 73-78
Keywords ALISA, Protein thiol, Biomarker, Redox signalling, Oxidative stress, Exercise, PP2A, Human
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