Organization of phosphatidylcholine and sphingomyelin in the surface monolayer of low density lipoprotein and lipoprotein(a) as determined by time-resolved fluorometry.
Fluorescent analogs of phosphatidylcholine (PC) and sphingomyelin (SM) labeled with diphenylhexatrienylpropionic acid (DPH) were prepared and incorporated into the surface layer of human low density lipoprotein (LDL) and lipoprotein(a) (Lp(a)). Fluorescence anisotropy measurements of DPH-PC and DPH-SM in both lipoprotein classes were carried out at different temperatures ranging from 20 to 37 degrees C. DPH-PC as well as DPH-SM were shown to reside in more rigid domains in Lp(a) than in LDL according to higher anisotropy values in Lp(a). In both LDL and Lp(a), DPH-PC experienced a more rigid environment than DPH-SM, suggesting different environments of PC and SM in the surface shell of the lipoproteins. Fluorescence lifetimes of the labeled lipoproteins were determined by phase and modulation fluorometry. We found bimodal Lorentzian distributions for the decay times of DPH-PC and DPH-SM in LDL and Lp(a). Lifetime distribution centers for labeled lipids were very similar except for DPH-PC in Lp(a) which was shifted to longer lifetimes, suggesting a less polar environment of PC in Lp(a) than in LDL. The distributional width of DPH-PC in Lp(a) was broader than in LDL. Accordingly, phosphatidylcholine must be localized in a more homogeneous environment in LDL as compared with Lp(a). On the other hand, no difference in distributional widths was observed for DPH-SM in both lipoproteins, showing that SM organization in Lp(a) is unaffected by apo(a). From the obtained fluorescence data we propose that apoproteins discriminate between the choline phospholipids and preferentially associate with phosphatidylcholine. This effect is enhanced in Lp(a) due to the presence of apolipoprotein(a).
Sommer
A
A
Institut für Biochemie und Lebensmittelchemie, Technische Universität Graz, Austria.
Prenner
E
E
Gorges
R
R
Stütz
H
H
Grillhofer
H
H
Kostner
G M
GM
Paltauf
F
F
Hermetter
A
A
D005454:Fluorescence Polarization; D006801:Humans; D017270:Lipoprotein(a) / Q000097:blood / Q000737:chemistry* / Q000302:isolation & purification; D008077:Lipoproteins, LDL / Q000097:blood / Q000737:chemistry* / Q000302:isolation & purification; D010713:Phosphatidylcholines / Q000032:analysis*; D010743:Phospholipids / Q000032:analysis*; D013050:Spectrometry, Fluorescence / Q000379:methods; D013109:Sphingomyelins / Q000032:analysis*; D013816:Thermodynamics; D013997:Time Factors
D017270:Lipoprotein(a); D008077:Lipoproteins, LDL; D010713:Phosphatidylcholines; D010743:Phospholipids; D013109:Sphingomyelins