Characterization of the discoidal complexes formed between apoA-I-CNBr fragments and phosphatidylcholine.
The structure, composition, and physico-chemical properties of lipid-protein complexes generated between dimyristoylphosphatidylcholine (DPMC) and the CNBr fragments of human apoA-I were studied. The fragments were separated by high performance liquid chromatography and purified on a reversed-phase column. The complexes with DMPC were isolated on a Superose column; their dimensions were obtained by gradient gel electrophoresis and by electron microscopy. The secondary structure of the protein in the complexes was studied both by circular dichroism and by attenuated total reflection infrared spectroscopy. The fragments 1 and 4 of apoA-I, containing, respectively, two and three amphipathic helices, recombined with the phospholipid to generate discoidal particles with sizes similar to that of apoA-I- and apoA-II-DMPC complexes. The infrared measurements indicated that in all complexes the apolipoprotein helical segments were oriented parallel to the phospholipid acyl chains and that the protein was located around the edges of the discs. Computer modelling of the complexes based on energy minimization techniques proposed a model for these particles in agreement with the dimensions measured experimentally. In conclusion, we propose that apoA-I and its longest CNBr fragments are able to generate discoidal particles with DMPC, with apolipoprotein helical segments oriented parallel to the acyl chains of the phospholipids.
Vanloo
B
B
Department of Clinical Chemistry, A.Z. St-Jan, Brugge, Belgium.
Morrison
J
J
Fidge
N
N
Lorent
G
G
Lins
L
L
Brasseur
R
R
Ruysschaert
J M
JM
Baert
J
J
Rosseneu
M
M
D016632:Apolipoprotein A-I; D001054:Apolipoproteins A / Q000737:chemistry*; D001665:Binding Sites; D002850:Chromatography, Gel; D003198:Computer Simulation; D003488:Cyanogen Bromide; D004134:Dimyristoylphosphatidylcholine / Q000737:chemistry*; D004789:Enzyme Activation; D005454:Fluorescence Polarization; D008854:Microscopy, Electron; D008958:Models, Molecular; D008968:Molecular Conformation; D009391:Nephelometry and Turbidimetry; D010455:Peptides / Q000737:chemistry*; D011489:Protein Denaturation; D013055:Spectrophotometry, Infrared
D016632:Apolipoprotein A-I; D001054:Apolipoproteins A; D010455:Peptides; D003488:Cyanogen Bromide; D004134:Dimyristoylphosphatidylcholine