Ubiquitin is a carbon dioxide-binding protein.

Linthwaite, Victoria L; Pawloski, Wes; Pegg, Hamish B; Townsend, Philip D; Thomas, Michael J; So, Victor K H; Brown, Adrian P; Hodgson, David R W et al. · Sci Adv · 2021

basic_science · Level V

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Abstract

The identification of CO<sub>2</sub>-binding proteins is crucial to understanding CO<sub>2</sub>-regulated molecular processes. CO<sub>2</sub> can form a reversible posttranslational modification through carbamylation of neutral N-terminal α-amino or lysine ε-amino groups. We have previously developed triethyloxonium (TEO) ion as a chemical proteomics tool for covalent trapping of carbamates, and here, we deploy TEO to identify ubiquitin as a mammalian CO<sub>2</sub>-binding protein. We use <sup>13</sup>C-NMR spectroscopy to demonstrate that CO<sub>2</sub> forms carbamates on the ubiquitin N terminus and ε-amino groups of lysines 6, 33, 48, and 63. We demonstrate that biologically relevant <i>p</i>CO<sub>2</sub> levels reduce ubiquitin conjugation at lysine-48 and down-regulate ubiquitin-dependent NF-κB pathway activation. Our results show that ubiquitin is a CO<sub>2</sub>-binding protein and demonstrates carbamylation as a viable mechanism by which mammalian cells can respond to fluctuating <i>p</i>CO<sub>2</sub>.