Figure 7.

Characterizing kinetic parameters of PHD3 with the ACC2 P450 peptide. A, PHD3 was added to 250 μM of the ACC2 P450 peptide and 0.5 mM α-ketoglutarate. For all negative controls, 0.5 mM a-ketoglutarate, 250 μM peptides, and 10 μM of PHD3 were used unless otherwise indicated. B, increasing concentrations of ACC2 peptide (P450) were added to saturating concentrations of all other reagents. C, relative abundance of detected peptide fragments. “b” fragments (blue) are N-terminal amino acid fragments of the peptide, and “y” fragments (red) are C-terminal amino acid fragments of the peptide. 250 μM of peptide was incubated with (left panel) or without (right panel) 10 μM PHD3 to generate the mass spectra above. Plotted data represent three independent replicates. α-KG, α-ketoglutarate.