Figure 6.

The formation of NAE from GP-NAE by GDE1. The radioactive band corresponding to GP-N-[¹⁴C]palmitoylethanolamine (GP-NPE), which was produced by purified cPLA₂γ, was scraped from the TLC plate. The radioactive compound was then extracted by the Bligh and Dyer protocol, and incubated with purified GDE1 (G1) or water (-) (A). N-[¹⁴C]Palmitoyl-PE (NPPE) was incubated with the purified cPLA₂γ for 30 min and additionally with the purified GDE1 for 15 min (γ+G1) (B). GDE1 or cPLA₂γ was omitted in γ and G1, respectively. The products were analyzed by TLC. The positions of the origin and authentic compounds are shown. The N-[¹⁴C]palmitoyl-lysoPE (lysoNPPE)-forming activity (C), GP-NPE-forming activity (D), and N-[¹⁴C]palmitoylethanolamine (NPE)-forming activity (E) are shown (mean values ± S.D., n = 3).