Experiments revealed that cytochrome P450 2C8 enzyme (CYP2C8) has two distinct substrate binding sites to the physiologically important molecules, retinoic acids, and the main difference between these two binding sites is whether there is a salt bridge interaction between the anionic carboxylate tail of retinoic acids and the surrounding protein environment. However, the influence of such salt bridge interaction toward catalysis is still elusive. In the present paper, density functional theory (DFT) calculations were employed to research the reaction mechanism of all-trans-retinoic acid (atRA) 4-hydroxylation mediated by CYP2C8. Our DFT calculations revealed that such salt bridge interaction has obvious effects on the reaction mechanism of atRA 4-hydroxylation. In the binding site containing a salt bridge interaction between the anionic carboxylate tail of atRA and the cationic guanidine group of Arg241, C – H bond activation proceeds via a normal hydrogen atom transfer (HAT) mechanism; in the other site without this salt bridge interaction, however, C – H bond activation is achieved via a stepwise electron transfer and hydrogen atom transfer, thus, a novel ET/HAT mechanism. These findings enrich the mechanism patterns of C – H bond activation catalyzed by metalloenzymes and their biomimetics. Meanwhile, the self-interaction error (SIE) problem encountered during our calculations in vacuum was affected and removed by the inclusion of an external electric field in the calculations.