Fig. 3
From: SIRT5 functions as a tumor suppressor in renal cell carcinoma by reversing the Warburg effect
SIRT5 desuccinylates PDHA1 at K351. (A) Lysine 77 (K77), K121, K374, K351 and 3 K (K77, K121 and K374) of PDHA1 were mutated to glutamine and introduced into HEK293T cells. Then, the pansuccinylation level of PDHA1 was determined in the presence or absence of SIRT5. B Validation of the specificity of the Succ-K351 antibody. Succ-K351 represented the peptide containing succinylated K351 and the flanking sequence. C Lysine 351 (K351) of PDHA1 was mutated to arginine (R) or glutamine (Q) and then transfected into HEK293T cells with or without SIRT5, and the pansuccinylation level of PDHA1 was determined. D K351R and K351Q PDHA1 mutants were transfected into HEK293T cells with or without SIRT5, and the succinylation level of PDHA1 was determined. E SIRT5∆50 was transfected into HeLa cells, and the succinylation level of endogenous PDHA1 was determined. F The succinylation levels of endogenous PDHA1 in HeLa cells before and after SIRT5 knockdown by independent shRNAs were compared