Biochemical and physical properties of remnant-HDL2 and of pre beta 1-HDL produced by hepatic lipase.
The hepatic lipase acting on triglyceride-rich high-density lipoprotein2 (HDL2) induces the formation of pre beta 1-HDL, leaving a residual alpha-migrating HDL particle that was named "remnant-HDL2" (Barrans, A., Collet, X., Barbaras, R., Jaspard, B., Manent, J., Vieu, C., Chap, H., and Perret, B. (1994) J. Biol. Chem. 269, 11572-11577.]. In this study, these two product particles generated by hepatic lipase were isolated by density gradient ultracentrifugation. Particles were first characterized in terms of chemical composition, density, and mass. The pre beta 1-HDL obtained in vitro contain one to two molecules of apoA-I, associated with phospholipids, and free and esterified cholesterol. When compared to triglyceride-rich HDL2, remnant-HDL2 have lost on average one molecule of apoA-I, 60% of triacylglycerols, and 15% of phospholipids. The estimated composition is concordant with the hypothesis of the splitting of a substrate particle into one pre beta 1-HDL and one remnant-HDL2. Spectroscopic studies were carried out to monitor changes in lipid fluidity upon lipolysis. The fluorescence anisotropy was measured using (1,6)-diphenyl-hexa-(1,3, 5)-triene as a probe, and the degree of order was calculated from electron spin resonance spectra using the 5-nitroxy-derivative of stearic acid. Both approaches showed a decreased lipid fluidity in remnant-HDL2, as compared to triglyceride-rich HDL2. The immunoreactivity of apoA-I toward several monoclonal antibodies was assayed as a reflection of changes of apoA-I conformation. In remnant-HDL2, as compared to triglyceride-rich HDL2, a lower reactivity was noted with the 2G11 antibody, which interacts in the NH2 terminal part of apoA-I. Finally, remnant-HDL2 was clearly different from HDL3 with respect to all of the parameters studied, demonstrating that hepatic lipase does not promote the direct conversion of HDL2 to HDL3. Thus, hepatic lipase produces remnant-HDL2 particles, which display modifications of apoA-I conformation and of fluidity of the lipid environment. This newly described HDL2 subfraction may play a major role in the reverse cholesterol transport.
Guendouzi
K
K
Institut National de la Santé et de la Recherche Médicale, Unité 326, Phospholipides membranaires, Signalisation cellulaire et Lipoprotéines, Hôpital Purpan, Toulouse, France.
Jaspard
B
B
Barbaras
R
R
Motta
C
C
Vieu
C
C
Marcel
Y
Y
Chap
H
H
Perret
B
B
Collet
X
X
D016632:Apolipoprotein A-I / Q000737:chemistry* / Q000276:immunology; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D004578:Electron Spin Resonance Spectroscopy; D005454:Fluorescence Polarization; D053442:High-Density Lipoproteins, Pre-beta; D006801:Humans; D015151:Immunoblotting; D008049:Lipase / Q000737:chemistry / Q000378:metabolism*; D008075:Lipoproteins, HDL / Q000096:biosynthesis* / Q000737:chemistry* / Q000302:isolation & purification; D008099:Liver / Q000201:enzymology*; D011487:Protein Conformation; D011863:Radioimmunoassay; D013050:Spectrometry, Fluorescence; D014461:Ultracentrifugation
D016632:Apolipoprotein A-I; D053442:High-Density Lipoproteins, Pre-beta; D008075:Lipoproteins, HDL; D008049:Lipase