Hydration and Localization of Diacylglycerol in the Insect Lipoprotein Lipophorin. A 13C-NMR Study

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Abstract
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.

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