Protein-L-isoaspartate O-methyltransferase is required for in vivo control of oxidative damage in red blood cells

• Published in: Haematologica (Roma) 2021-10, Vol.106 (10), p.2726-2739
• Main Authors: D'Alessandro, Angelo, Hay, Ariel, Dzieciatkowska, Monika, Brown, Benjamin C, Morrison, Evan J, Hansen, Kirk C, Zimring, James C
• Format: Article
• Language: English
• Subjects: Animals; Erythrocytes - metabolism; Isoaspartic Acid - metabolism; Oxidative Stress; Protein D-Aspartate-L-Isoaspartate Methyltransferase - genetics; Protein D-Aspartate-L-Isoaspartate Methyltransferase - metabolism; Reactive Oxygen Species
• ISSN: 0390-6078; 1592-8721
• DOI: 10.3324/haematol.2020.266676

Red blood cells have the special challenge of a large amount of reactive oxygen species (from their substantial iron load and Fenton reactions) combined with the inability to synthesize new gene products. Considerable progress has been made in elucidating the multiple pathways by which red blood cells neutralize reactive oxygen species via NADPH driven redox reactions. However, far less is known about how red blood cells repair the inevitable damage that does occur when reactive oxygen species break through anti-oxidant defenses. When structural and functional proteins become oxidized, the only remedy available to red blood cells is direct repair of the damaged molecules, as red blood cells cannot synthesize new proteins. Amongst the most common amino acid targets of oxidative damage is the conversion of asparagine and aspartate side chains into a succinimidyl group through deamidation or dehydration, respectively. Red blood cells express an L-Isoaspartyl methyltransferase (PIMT, gene name PCMT1) that can convert succinimidyl groups back to an aspartate. Herein, we report that deletion of PCMT1 significantly alters red blood cell metabolism in a healthy state, but does not impair the circulatory lifespan of red blood cells. Through a combination of genetic ablation, bone marrow transplantation and oxidant stimulation with phenylhydrazine in vivo or blood storage ex vivo, we use omics approaches to show that, when animals are exposed to oxidative stress, red blood cells from PCMT1 knockout undergo significant metabolic reprogramming and increased hemolysis. This is the first report of an essential role of PCMT1 for normal RBC circulation during oxidative stress.