Conformation of apolipoprotein AI in reconstituted lipoprotein particles and particle-membrane interaction: effect of cholesterol.
Discoidal recombinant high density lipoproteins (rHDL) of apolipoprotein AI (apoAI) and palmitoyloleoylphosphatidylcholine (POPC), with or without cholesterol, were prepared by cholate dialysis. By gel filtration, rHDL containing 2-4 (Lp2, Lp3 and Lp4) apoAI molecules/particle were obtained. The ApoAI conformation in these rHDL was investigated by tryptophan fluorescence, denaturation with guanidine HCl, and immunoreactivity with two monoclonal antibodies recognizing epitopes in the N-terminal and central domains. Data show that apoAI conformation is highly dependent on particle size as well as on cholesterol. The ability of rHDL to interact with lipid bilayer was studied by measuring leakage induction on POPC and POPC/cholesterol vesicles loaded with terbium/dipicolinic acid. Among the cholesterol-free rHDL, the most efficient ones were the smallest Lp2. Leakage induction on POPC vesicles is dramatically decreased by the presence of cholesterol in Lp2 and Lp3. All the rHDL, but specially those containing cholesterol, induced more leakage on the POPC/cholesterol than on the POPC vesicles. These results suggest that in small cholesterol-poor particles, apoAI could have a conformation determining a high affinity for membranes, which could facilitate cholesterol efflux. After cholesterol enrichment, a conformational change in apoAI could decrease the affinity for membranes allowing the lipoprotein release.
Tricerri
A
A
Instituto de Investigaciones Bioquímicas de La Plata (INIBIOLP), CONICET-UNLP, Facultad de Ciencias Médicas, calles 60 y 120, 1900-La Plata, Argentina.
Córsico
B
B
Toledo
J D
JD
Garda
H A
HA
Brenner
R R
RR
D000911:Antibodies, Monoclonal / Q000276:immunology / Q000378:metabolism; D016632:Apolipoprotein A-I / Q000737:chemistry* / Q000276:immunology; D001667:Binding, Competitive; D002784:Cholesterol / Q000494:pharmacology*; D000939:Epitopes / Q000737:chemistry / Q000276:immunology; D005453:Fluorescence; D019791:Guanidine / Q000494:pharmacology; D008075:Lipoproteins, HDL / Q000737:chemistry / Q000235:genetics; D008081:Liposomes / Q000378:metabolism*; D010316:Particle Size; D010539:Permeability; D010713:Phosphatidylcholines / Q000378:metabolism; D010848:Picolinic Acids / Q000378:metabolism; D011487:Protein Conformation; D011489:Protein Denaturation / Q000187:drug effects; D013725:Terbium / Q000378:metabolism; D014364:Tryptophan / Q000378:metabolism
D000911:Antibodies, Monoclonal; D016632:Apolipoprotein A-I; D000939:Epitopes; D008075:Lipoproteins, HDL; D008081:Liposomes; D010713:Phosphatidylcholines; D010848:Picolinic Acids; D013725:Terbium; C067490:terbium chloride; D014364:Tryptophan; D002784:Cholesterol; D019791:Guanidine; C028694:1-palmitoyl-2-oleoylphosphatidylcholine; C004860:dipicolinic acid