Circular dichroic spectra of apolipoprotein E in model complexes and cholesterol-rich lipoproteins: lipid contribution

• Published in: Biochemistry (Easton) 1984-12, Vol.23 (26), p.6530-6538
• Main Authors: Chen, G. Chi, Guo, Luke S. S, Hamilton, Robert L, Gordon, Virginia, Richards, E. Glen, Kane, John P
• Format: Article
• Language: English
• Subjects: Animals; Apolipoproteins E; Cholesterol, Dietary - administration & dosage; Circular Dichroism; Dogs; Lipids; Lipoproteins, HDL; Liposomes; Protein Conformation
• ISSN: 0006-2960; 1520-4995
• DOI: 10.1021/bi00321a039

Lipid-free apolipoprotein E (apo E) and canine apo E HDLc, a cholesterol-rich lipoprotein containing apo E as the only apolipoprotein, show very different circular dichroism (CD) spectra. To determine the cause of the spectral difference, we estimated the CD contribution of phospholipid, cholesterol, and cholesteryl ester in liposomes and microemulsions. We prepared microemulsions, containing nearly equal amounts of egg phosphatidylcholine (PC) and cholesteryl oleate (mean diameter 320 A), by an injection technique. Both microemulsions and cholesterol-containing liposomes exhibit intense negative CD bands in the far-ultraviolet region. Lipids contribute about 20% of the spectral difference between apo E and apo E HDLc at 222 nm, and about 60% of the spectral difference at 208 nm. The remainder of the spectral difference is attributable to lipid-protein interaction corresponding to a 15-30% increase in helicity of apo E. CD analysis indicates that the helical content of apo E in apo E HDLc resembles that in the ternary complex apo E-PC-cholesterol (or apo E-PC-cholesteryl ester) more than that in the binary complex apo E-PC, suggesting that cholesterol affects the conformation of apo E. Our data indicate that in going from a lipid-free state to a lipid environment, apo E undergoes a random to helix transition, assuming the maximal helicity predicted from its primary structure.