Uncovering the Biosynthetic Pathway of Oleamide

Oleamide belongs to a recently discovered class of signaling molecules called primary fatty acid amides (PFAMs). As of yet, the in vivo biosynthetic pathway for oleamide and the other cell‐signalling PFAMs is unknown. The pathway we have proposed is shown in Fig. 1. 1Hypothesized Pathway for PFAM Biosynthesis RT‐PCR and Western analysis were employed to determine which enzymes from the pathway shown in Fig. 1 are expressed in mouse neuroblastoma N 18 TG 2 cells (which are known to produce oleamide from oleic acid), choroid plexus cells, liver cells, and kidney cells. The expression of ACS and PAM in the N 18 TG 2 cells supports the proposed biosynthetic pathway. None of the enzymes thought to catalyze reaction 2 in Fig. 1 (bile acid:amino acid transferase, acyl‐CoA:glycine N ‐acyltransferase, or N ‐myristoyl transferase) were expressed in N 18 TG 2 cells. This suggests that an undiscovered enzyme exists for the biosynthesis of the N ‐fatty acylglycine intermediates. Previous work from the Merkler laboratory has shown that addition of a PAM inhibitor to the N 18 TG 2 cells culture medium results in accumulation of N ‐oleoylglycine. Future directions include identifying the enzyme responsible for production of the N ‐fatty acylglycines and using RNAi to target the PFAM biosynthetic enzymes to examine the accumulation of metabolic precursors.