Skip to main content
. 2016 Mar 9;11(3):e0151085. doi: 10.1371/journal.pone.0151085

Fig 3. Validation of RBL-2_410–422 O-GlcNAcylation.

Fig 3

A, O-GlcNAcylation of RBL-2_410–422 peptide by bacterial lysate containing m-OGT (7 μg/μL) with increasing concentration of UDP-GlcNAc (0–10 mM). B, O-GlcNAcylation of RBL-2_410–422 peptide dependency of increasing total protein concentrations of bacterial lysate containing m-OGT (0, 1.7, 3.5, 7 μg/μL) at 1 mM UDP-GlcNAc. C, O-GlcNAcylation of RBL-2_410–422 peptide by purified m-OGT (0.2 μg/μL) with 1 mM UDP-GlcNAc was inhibited by a known OGT inhibitor (ST045849, 0–200 μM). D, Km value for UDP-GlcNAc was determined with fixed saturating concentration of RBL-2_410–422 peptide, purified m-OGT (0.2 μg/μL) and varying concentration of UDP-GlcNAc (0–2 mM). The Km value derived from the fit to Michaelis-Menten model is 24 μM.