Effect of particle size and temperature on the conformation and physiological behavior of apolipoprotein E bound to model lipoprotein particles.
The effect of particle size and structural order/disorder of the lipid domain on the conformation and physiological behavior of lipid-associated apolipoprotein E (apoE) was evaluated. Circular dichroic (CD) spectra of apoE bound to large (LME) and small (SME) microemulsion particles, composed of dimyristoylphosphatidylcholine (DMPC) and cholesteryl oleate (CO), and to DMPC disks revealed that at 4 degrees C, where all of the lipid constituents were in an ordered state, apoE bound to LME displayed approximately 60% alpha-helicity, while apoE bound to SME and DMPC disks displayed 73% and 95% helicity, respectively. Over the temperature range 4-50 degrees C, encompassing the lipid thermal transitions, only apoE bound to LME demonstrated an abrupt change in its CD spectrum (decrease in alpha-helicity) in response to temperature. To determine the source of the abrupt CD change, the constants for dissociation (Kd) of apoE from the surface of the large and small microemulsion particles were determined at 4, 25, and 37 degrees C. These results demonstrated that at 4 degrees C, the KdS for binding of apoE to the LME and SME were approximately equal; however, between 4 and 25 degrees C, there was a 5-fold increase in the Kd for binding of apoE to the LME, whereas the Kd for binding to the SME remained constant. The physiological effects of these differences in apoE secondary structure and equilibrium binding were examined by measuring the capacity of each apoE-containing particle to complete with LDL for binding to human fibroblasts, and by measuring the capacity of the apoE-microemulsion particles to suppress HMG-CoA reductase activity.(ABSTRACT TRUNCATED AT 250 WORDS)
Mims
M P
MP
Department of Medicine, Baylor College of Medicine, Houston, Texas.
Soma
M R
MR
Morrisett
J D
JD
D001057:Apolipoproteins E / Q000378:metabolism* / Q000648:ultrastructure; D001692:Biological Transport; D002788:Cholesterol Esters / Q000378:metabolism; D002942:Circular Dichroism; D004134:Dimyristoylphosphatidylcholine / Q000378:metabolism; D004655:Emulsions; D005347:Fibroblasts / Q000378:metabolism; D006801:Humans; D019161:Hydroxymethylglutaryl-CoA Reductase Inhibitors; D008074:Lipoproteins / Q000378:metabolism* / Q000648:ultrastructure; D010316:Particle Size; D011485:Protein Binding; D011487:Protein Conformation; D011956:Receptors, Cell Surface / Q000378:metabolism; D018110:Receptors, Lipoprotein; D013696:Temperature
D001057:Apolipoproteins E; D002788:Cholesterol Esters; D004655:Emulsions; D019161:Hydroxymethylglutaryl-CoA Reductase Inhibitors; D008074:Lipoproteins; D011956:Receptors, Cell Surface; D018110:Receptors, Lipoprotein; C014409:cholesteryl oleate; D004134:Dimyristoylphosphatidylcholine