Fig. 5: AKT1-R391 methylation promotes AKT conformational change, membrane localization, and interaction with PDK1.
From: PRMT5-mediated arginine methylation activates AKT kinase to govern tumorigenesis
a IB analysis of WCLs and GST pulldown products derived from HEK293 cells transfected with indicated constructs. b Biotin-conjugated R391 peptides with (me2s) or without (me0) dimethylation were incubated with recombinant GST-AKT1-PH and precipitated with streptavidin. Peptide (4 μg) was used for each sample. c IB analyses of PIP3 pulldown products. HA-AKT1 proteins were purified from HEK293T cells transfected with HA-AKT1-WT or HA-AKT1-R391K. Empty beads (ctr) serve as a negative control. d IB analyses of PIP3 pulldown products and WCLs derived from PRMT5-knockout MCF7 cells. e, f IB analysis of cell fractionations derived from PRMT5-depleted MCF7 cells (e) or from DLD-1-AKT1/2−/− cells expressing AKT1-WT or R391K mutant (f). g IB analysis of WCLs and IP products derived from PRMT5-knockdown MCF7 cells. h IB analysis of WCLs and IP products derived from DLD-1-AKT1/2−/− cells expressing AKT1-WT or R391K mutant. Cells were serum-starved for 16 h and then treated with 100 nM insulin for 60 min before collection. Similar results were obtained in n ≥ 2 independent experiments in a–h. Uncropped immunoblots are provided in Source Data file.
