FIGURE 1.

Workflow for investigating SIRT5-induced malonylome and succinylome remodeling in mouse brain. Biological quadruplicates of wild-type and Sirt5^(−/−) mouse brains were prepared for in solution trypsin digestion. Samples were enriched for malonyl (Ma) peptides or succinyl (Suc) peptides using antibody-conjugated beads. PTM-enriched samples and samples resulting from whole-lysate digestion were individually analyzed on a nanoLC-Orbitrap Eclipse Tribrid system operating in DIA mode. Finally, data were processed with directDIA (Biognosys) using a library-free strategy for identification, quantification, and PTM site localization. DIA, data-independent acquisition; PTM, post-translational modification; SIRT5, Sirtuin 5