Figure 2. NRMT can bind to its fully methylated substrate proteins.
A) NRMT is co-precipitated with C-terminally FLAG-tagged substrates (RCC1 and VRK2)
B) C-terminally FLAG-tagged NRMT co-precipitates with endogenous ZFP15 and Mina53. IPs were run on the 10% polyacrylamide gel and stained with Commassie blue.
C) C-terminally FLAG-tagged wild type ZFP15 co-precipitates with endogenous NRMT. The PPQ non-methylatable mutant of ZFP15 does not bind endogenous NRMT.
D) Both PPK- and PPQ-ZFP15 FLAG-tagged proteins are efficiently expressed and immunoprecipitated.
E) N-terminal methylation activity is localized in the nucleus. NLS fused SPK-GFP3 substrates are efficiently methylated, in contrast to NES fused SPK-GFP3.