FIGURE 3.

C-terminal tail of PGAM1 is acetylated. A, MYC-PGAM1 and MYC-PGAM1 3KQ mutant (K251Q, K253Q, and K254Q) were transiently transfected into HEK293 cells following 5 mm NAM treated for 16 h. Cell lysates were immunoprecipitated (IP) with anti-MYC antibody, resolved by SDS-PAGE, and then detected with Western blot (WB) analysis with anti-acetyl-lysine (AcK) and anti-MYC antibodies. B and C, activity was measured at 340 nm by the PGAM1-coupled assay. D, PGAM1 3KQ shows high catalytic activity against 2-phosphoglycerate. Catalytic amounts of PGAM1 ●, PGAM1 3KQ ■, and PGAM1 3KR ⧫ were incubated with the indicated amounts of 3-PGA, and activity was measured at 340 nm by PGAM1-coupled assay. Data from the coupled assay were fitted to the Michaelis-Menten equation. Error bars represent S.E. (n = 3). *, p < 0.05.