Role of glycosylation in the lipid-binding activity of the exchangeable apolipoprotein, apolipophorin-III

Non-glycosylated recombinant Locusta migratoria apolipophorin-III, apoLp-III, was expressed in E. coli and its physical-chemical properties were compared to those of the glycosylated native apoLp-III. Fluorescence quantum yield and acrylamide quenching studies indicated a slightly higher accessibility of the Trp residues in the recombinant apoLp-III. Far-UV CD spectroscopy indicated that the recombinant apoLp-III has a lower alpha-helical content than the glycosylated apoLp-III. Both proteins spontaneously formed discoidal recombinant lipoprotein particles when incubated with dimyristoylphosphatidylcholine (DMPC). Interaction with lipid promotes an increase in a-helical content. CD and fluorescence studies indicate that both proteins adopt the same conformation in the lipid-bound state. However, the kinetics of association of the recombinant protein with DMPC is 5-fold faster than that of the native protein. The results suggest that glycosylation inhibits the lipid binding activity by preventing the exposure of hydrophobic domains and/or decreasing the conformational flexibility of the protein. (C) 1998 Academic Press.