Fig. 3: Lysine 89 acetylation promotes partial unwinding of helix αb’ and ubiquitylation of residues K95/97.
From: Acetylation-dependent coupling between G6PD activity and apoptotic signaling
a Lysine 89 acetylation promotes ubiquitylation of G6PD. Representative Western blots showing the ubiquitylation level of immunoprecipitated Flag-tagged G6PD variants, co-expressed with HA-tagged ubiquitin in HEK293T cells treated with 5 μM MG132 for 16 h. b Mutation of lysine 95 and 97 to arginine does not affect G6PD activity. Bars represent the relative Vmax of pWT, AcK89, and indicated Lys-to-Arg G6PD mutants, measured in lysates of cells incubated in the absence of KDACi, and displayed relative to pWT G6PD. Data are the mean ± SD, n = 3 biologically independent samples. c AcK89 G6PD is ubquitylated on residues K95/97. Representative Western blots showing the ubiquitylation level of immunoprecipitated Flag-tagged G6PD variants, co-expressed with HA-tagged ubiquitin in HEK293T cells treated with 5 μM MG132 for 16 h. d Lysine 89 acetylation has minimal effect on the three-dimensional structure of monomeric G6PD. Superposition of AcK89 G6PD (green) and non-acetylated G6PD (cyan, PDB ID: 6E08). Structural NADP+ and K89 are shown in sticks model. e Lysine 89 acetylation promotes local conformational changes. Left: zoom in on K89 and nearby helices, showing the position of AcK89 (green) relative to the position of K89 (cyan, PDB ID: 6E08). Right: electron density map around residue AcK89. The 2Fo–Fc map was contoured at 1 σ. f Lysine 89 acetylation promotes partial unwinding of helix αb'. Local conformational changes around AcK89 with the partial unwinding of helix αb' (green), relative to the structure of non-acetylated G6PD (cyan, PDB ID: 6E08). Source data are provided as a Source Data file.
