The endogenous cannabinoid ligand, 2-arachidonoylglycerol (2-AG), is a lipid-derived signaling molecule that activates cannabinoid receptors in brain and peripheral tissues. Because of high lipophilicity, 2-AG readily partitions into lipid bilayers. Molecular behavior of 2-AG in bilayers and mechanism of approach to the receptor binding-pocket remain unsolved; approach via lateral diffusion in lipid matrix was suggested according to microsecond-long molecular dynamics simulations. We studied location, structure, dynamics, and kinetics of acyl-chain migration (2-AG→1-AG) of 2-AG in lipid bilayers of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) by solid-state NMR and neutron diffraction. Binding of 2-AG to purified CB2 receptor functionally reconstituted into liposomes was probed by 2H magic-angle-spinning NMR of headgroup-deuterated 2-AG (2-AG-d5). The exponential-decay constant of 2-AG due to acyl-chain migration at physiological temperature was determined to be 43 min, while molecular characterization of 2-AG was achieved by investigating the temperature-dependence. 1H cross-relaxation rates measured in two-dimensional NOESY revealed that the double-bond region of the arachidonoyl chain (20:4) of 2-AG locates with high probabilities at the upper methylene segments of POPC, with a significant distribution around glycerol. The data agrees with neutron diffraction, exhibiting highest probabilities of those double bonds near the hydrophobic-hydrophilic interface. The arachidonoyl chain shows high flexibility in lipid bilayers. In the presence of CB2, 2H resonances of 2-AG-d5 shifted upfield with line-broadening. The upfield shift was absent when the pocket was blocked with a high affinity agonist like CP-55,940, demonstrating specificity of 2-AG interaction with the CB2 binding-pocket. The induced shift suggests a model of rapid exchange of 2-AG between free and CB2-bound states; discussions regarding other possibilities will be presented. The enrichment of 2-AG in lipid bilayers and the conformational flexibility of 2-AG are likely to be critical for an efficient interaction with CB2.