Evidence that apolipoprotein A-I facilitates hepatic lipase-mediated phospholipid hydrolysis in reconstituted HDL containing apolipoprotein A-II

This study examines hepatic lipase (HL) mediated phospholipid hydrolysis in mixtures of apolipoprotein-specific, spherical reconstituted high-density lipoproteins (rHDL). We have shown previously that apolipoprotein A-I (apoA -I) and apoA-II have a major influence on the kinetics of HL-mediated phosp holipid and triacylglycerol hydrolysis in well-characterized, homogeneous p reparations of spherical rHDL [Hime, N. J., Barter, P. J., and Rye, K.-A. ( 1998) J. Biol. Chem. 273, 27191-27198]. In the present study, phospholipid hydrolysis was assessed in mixtures of rHDL containing either apoA-I only, (A-I)rHDL, apoA-II only, (A-II)rHDL, or both apoA-I and apoA-II, (A-YA-II)r HDL. The rHDL contained trace amounts of radiolabeled phospholipid, and hyd rolysis was measured as the formation of radiolabeled nonesterified fatty a cids (NEFA). As predicted from our previous kinetic studies, the (A-II)rHDL acted as competitive inhibitors of HL-mediated phospholipid hydrolysis in (A-I)rHDL. Less expected was the observation that the rate of phospholipid hydrolysis in (A II)rHDL was enhanced when (A-I)rHDL were also present in t he incubation mixture. The rate of phospholipid hydrolysis in (A-YA-II)rHDL was also greater than in (A-II)rHDL, indicating that apoA I enhances phosp holipid hydrolysis when it is present as a component of (A-I/A-II)rHDL. It is concluded that apoA I enhances HL-mediated phospholipid hydrolysis in ap oA-II containing rHDL, irrespective of whether the apoA-I is present in the same particle as the apoA-II [as in (A-I/A-II)rHDL] or whether it is prese nt as a component of a different particle, such as when (A-I)rHDL are added to incubations of (A-II)rHDL.