In the most stable binding pose of LysoPS(18:1) to GPR34 (Figure ), the dihedral angles corresponding to D1, D2 and CV1, CV2 of the glycerol moiety are +99.8˚(corresponding to D1 and CV1) and -155.2˚(corresponding to D2 and CV2) (Figure ). These dihedral angles are consistent with both the distributions provided by REMD (Figure ; yellow dashed circle) and the stable energy states obtained by MTD (Figure ; black dashed circle). However, it should be noted that short-duration REMD did not detect the minor conformer of LysoPS(18:1) that finally turned out to be the bioactive conformer. Nevertheless, the results suggested that the bioactive conformation of LysoPS(18:1) bound to GPR34 is present among the unbound ligand conformations. Intriguingly, this bioactive conformation is located in a minor distribution region, but would be selected upon binding to GPR34. The energy penalty to take a minor conformation, that is, the energy difference between the energy minima conformation (G=0.00 kcal/mol in MTD, e. g. CV1=177.46°, CV2=-172.87°) and the conformation in MTD approximately corresponding to the binding pose of LysoPS(18:1) (CV1=99.66°, CV2=-155.59°) is 4.26 kcal/mol, which will be compensated by binding to GPR34.