Hydration and localization of diacylglycerol in the insect lipoprotein lipophorin. A 13C-NMR study.
In order to probe the organization of diacylglycerol (DG) in lipophorin, 13C-enriched lipophorin was prepared for NMR investigations. We obtained 13C-enriched lipophorin labeled exclusively in DG by feeding insects tobacco leaves coated with [1-13C]palmitic acid or [1-13C]oleic acid. Lipophorins enriched up to 5% with a [13C]fatty acid were obtained by this procedure. NMR studies of the isolated lipophorin DG showed that palmitic acid accumulates almost entirely (> 90%) in the sn-1 position. Oleic acid was found equally distributed between the sn-1 and sn-2 positions, yielding a DG enriched equally at both positions. The 13C-NMR spectra of both [13C]palmitate- and [13C]oleate-enriched lipophorins showed that DG had one narrow carbonyl resonance indicative of rapid motion. A comparative analysis of the 13C carbonyl chemical shift data for DG in organic solvents, aqueous solutions, and dispersions with the DG carbonyl chemical shift of native lipophorin enriched in [13C]palmitate or [13C]oleate shows a high degree of water exclusion from the DG carbonyls in lipophorin. This result is consistent with the existence of a lipophorin lipid core containing most of the lipophorin DG. This study represents the first attempt to elucidate the organization of DG in lipophorin. The possibility of obtaining [13C]DG-enriched lipophorins, selectively enriched in one or both acyl chains of DG, should provide a powerful tool for further analysis of the organization and the dynamic properties of DG in native lipoproteins.
Soulages
J L
JL
Department of Biochemistry, University of Arizona, Tucson 85721.
Rivera
M
M
Walker
F A
FA
Wells
M A
MA
D000818:Animals; D002247:Carbon Isotopes; D002352:Carrier Proteins / Q000737:chemistry* / Q000378:metabolism; D004075:Diglycerides / Q000032:analysis / Q000737:chemistry*; D004121:Dimethyl Sulfoxide; D008055:Lipids / Q000737:chemistry*; D008074:Lipoproteins; D009682:Magnetic Resonance Spectroscopy; D009036:Moths; D019301:Oleic Acid; D009829:Oleic Acids / Q000378:metabolism; D019308:Palmitic Acid; D010169:Palmitic Acids / Q000378:metabolism; D014867:Water
D002247:Carbon Isotopes; D002352:Carrier Proteins; D004075:Diglycerides; D008055:Lipids; D008074:Lipoproteins; D009829:Oleic Acids; D010169:Palmitic Acids; C032830:lipophorin; D014867:Water; D019301:Oleic Acid; D019308:Palmitic Acid; D004121:Dimethyl Sulfoxide