Kr. Kozak et al., Amino acid determinants in cyclooxygenase-2 oxygenation of the endocannabinoid 2-arachidonylglycerol, J BIOL CHEM, 276(32), 2001, pp. 30072-30077
The endocannabinoid, 2-arachiclonylglycerol (2-AG), is an endogenous ligand
for the central (CBI) and peripheral (CB2) cannabinoid receptors and has b
een shown to be efficiently and selectively oxygenated by cyclooxygenase (C
OX)-2. We have investigated 2-AG/COX-2 interactions through site-directed m
utagenesis. An evaluation of more than 20 site-directed mutants of murine C
OX-2 has allowed for the development of a model of 2-AG binding within the
COX-2 active site. Most strikingly, these studies have identified Arg-513 a
s a critical determinant in the ability of COX-2 to efficiently generate pr
ostaglandin Hz glycerol ester, explaining, in part, the observed isoform se
lectivity for this substrate. Mutational analysis of Leu-531, an amino acid
located directly across from Arg-513 in the COX-2 active site, suggests th
at 2-AG is shifted in the active site away from this hydrophobic residue an
d toward Arg-513 relative to arachidonic acid. Despite this difference, asp
irin-treated COX-2 oxygenates 2-AG to afford 15-hydroxyeicosatetraenoic aci
d glycerol ester in a reaction analogous to the C-15 oxygenation of arachid
onic acid observed with acetylated COX-2. Finally, the differences in subst
rate binding do not alter the stereospecificity of the cyclooxygenase react
ion; 2-AG-derived and arachidonic acid-derived products share identical ste
reochemistry.