Interaction of CETP inhibitory peptide and lipoprotein substrates in cholesteryl ester transfer assay: relationship between association properties and inhibitory activities

In a previous study, CETP inhibitory peptide (3 kDa) was isolated from hog plasma. The peptide, synthesized chemically according to the amino acid sequence of the 3-kDa peptide (designated P₂₈), showed CETP inhibitory activity both in vitro and in vivo Cho et al. (1998) Biochim. Biophys. Acta 1391, 133–144]. We report herein further unique features of P₂₈ when it was associated with the cholesteryl ester (CE)-donor and-acceptor lipoproteins. Lipoprotein substrates with P₂₈ present in both HDL (as a CE-donor) and LDL (as a CE-acceptor) served as poor substrates, with CE-transfer activity decreased up to 60% compared to normal substrates without P₂₈. P₂₈ was found to be located in HDL fractions of hog plasma and showed the same electromobility as that visualized by PAGE on 7% polyacrylamide gel under nondenaturing conditions. Addition of apolipoprotein A-1 (apoA-1) or apoB antibody to a normal CE-transfer mixture did not alter CE-transfer activity. However, addition of apoA-1 or −B antibody to a CETP-inhibition mixture decreased the inhibitory activity of P₂₈ by ca. 20%. Western blot analysis revealed that P₂₈ was associated only with human and hog HDL among several lipoproteins purified from human, hog, and rabbit. CFTP-inhibition assays with various lipoprotein substrates revealed that P₂₈ exhibited substrate-specific inhibitory activity. The inhibitory activity of P₂₈ was highly dependent on the type of lipoprotein substrate (whether CE-donor or-acceptor); P₂₈ inhibited CE transfer from HDL to LDL, but it did not inhibit CE transfer from HDL to HDL.