FIGURE 4.
Expression and GP-NAE phosphodiesterase activity of GDE domain containing enzymes. A, Myc-tagged GDE-domain containing enzymes were overexpressed in COS-7 cells. Membrane fractions were treated with PNGase F and visualized by SDS-PAGE and Western blotting with an anti-Myc antibody (Cell Signaling). B, GP-NAE phosphodiesterase activity of membrane fractions of COS-7 cells expressing GDE domain-containing proteins was measured by monitoring release of N-C16:0 NAE via LC-MS with 100 μm N-C16:0 GP-NAE as substrate.