The exact biosynthesis of NaGly is not completely understood, but there are two proposed pathways found in vitro for its biosynthesis: 1) enzymatically regulated conjugation of arachidonic acid and glycine and 2) the oxidative metabolism of the endogenous cannabinoid anandamide. In the first pathway, Cytochrome c catalyzes the in vitro synthesis of NaGly from arachidonoyl coenzyme A and glycine in the presence of hydrogen peroxide. In the second pathway, alcohol dehydrogenase catalyzes the oxidation of anandamide into N-arachidonoyl glycine.
Effects on the nervous system
NAGly has been hypothesized to have a neurophysiological function of pain suppression, supported by evidence that it suppresses formalin-induced pain behavior in rats. In particular, peripherally administered NAGly inhibited phase 2 pain behavior, suggesting either a direct suppression of nociceptive afferents on the nerve or an indirect modulation of the afferents' interstitial environment. In either case, these findings hold promise for NAGly as a means of mitigating postoperative or chronic pain. NAGly is also effective in acute pain models, reducing mechanical allodynia and thermal hyperalgesia induced by intraplantar injection of Fruend's complete adjuvant. Similar mechanical allydonia induced by partial ligation of the sciatic nerve was also reduced by NaGly. Other arachidonic acid-amino acid conjugates did not have the same effects and the actions of NaGly were not affected by cannabinoid receptor agonists in either study, suggesting a novel non-cannabinoid receptor mediated approach to alleviate inflammatory pain.
NaGly was shown to be endogenous ligand for the G-protein couple receptor GPR92 along with farnesyl pyrophosphate. In the dorsal root ganglia (DRG), where GPR92 was found to be localized NaGly increased intracellular calcium levels in DRG neurons, indicating a role of NaGly in the sensory nervous system through the activation of GPR92.
Effects on the immune system
NAGly has been the focus of research on the immune system because of its antinociceptive effects and inhibitory action on components of the immune system. Specifically, it significantly inhibited TNFα and IFNγ production, and it shows potential as a therapeutic treatment for chronic inflammation. Moreover, NAGly has been shown to act as a substrate for cyclooxygenase-2 (COX-2), the enzyme primarily known for producing prostaglandins associated with increases in inflammation and hyperalgesia. In many mammalian tissues that express COX-2, significant levels of NAGly are naturally present, and in these tissues COX-2 selectively metabolizes NAGly prostaglandin (PG) H2 glycine and HETE-Gly.
NAGly has been hypothesized to induce cell migration in BV-2 microglia cells. The same research suggests that this migration occurs through GPR18. This was verified using GPR18 transfected HEK-293 cells. The same migration wasn't witnessed using non-transfected and GPR55 transfected HEK-293. Additionally, tetrahydrocannabinol and NaGly are full agonists at the GPR18 receptors and induce migration in human endometrial HEC-1B cells. Understanding functions of NaGly in such structures provides a promising future in helping treat diseases such as endometriosis.
NaGly was identified as a novel insulinsecretagogue and was shown to increase intracellular calcium concentration through stimulation of voltage dependent calcium channels. Additionally, this action was dependent on extracellular glucose level.
Additional biochemical interactions
NaGly has been shown to inhibit the glycine transporter GLYT2a in a non-competitive fashion with arachidonic acids and secondary messenger systems of GLYT2a, suggesting a novel recognition site for the N-arachodnoyl amino acids, especially because other conjugated amino acids had similar effects.
^Kohno M, Hasegawa H, Inoue A, Muraoka M, Miyazaki T, Oka K, Yasukawa M (September 2006). "Identification of N-arachidonylglycine as the endogenous ligand for orphan G-protein-coupled receptor GPR18". Biochemical and Biophysical Research Communications. 347 (3): 827–32. doi:10.1016/j.bbrc.2006.06.175. PMID16844083.
^McCue JM, Driscoll WJ, Mueller GP (January 2008). "Cytochrome c catalyzes the in vitro synthesis of arachidonoyl glycine". Biochemical and Biophysical Research Communications. 365 (2): 322–7. doi:10.1016/j.bbrc.2007.10.175. PMID17986381.
^ abHuang SM, Bisogno T, Petros TJ, Chang SY, Zavitsanos PA, Zipkin RE, Sivakumar R, Coop A, Maeda DY, De Petrocellis L, Burstein S, Di Marzo V, Walker JM (November 2001). "Identification of a new class of molecules, the arachidonyl amino acids, and characterization of one member that inhibits pain". The Journal of Biological Chemistry. 276 (46): 42639–44. doi:10.1074/jbc.M107351200. PMID11518719.
^WO application 9738688, Ferrante A, Poulos A, Pitt M, Easton C, Sleigh M, Rathjen D, Widmer F, "Methods of Treating Immunopathologies Using Polyunsaturated Fatty Acids", published 23 October 1997, assigned to Peptide Technology Pty Ltd. and Women's and Children's Hospital Adelaide
^Prusakiewicz JJ, Kingsley PJ, Kozak KR, Marnett LJ (August 2002). "Selective oxygenation of N-arachidonylglycine by cyclooxygenase-2". Biochemical and Biophysical Research Communications. 296 (3): 612–7. doi:10.1016/s0006-291x(02)00915-4. PMID12176025.