Investigation of human low-density lipoprotein by 1H nuclear magnetic resonance spectroscopy: Mobility of phosphatidylcholine and sphingomyelin headgroups characterizes the surface layer

H. C. Murphy, S. P. Burns, J. J. White, Jimmy D. Bell, R. A. Iles

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10 Citations (Scopus)

Abstract

The resolution of the trimethyl headgroup resonance of phosphatidylcholine (PC) and sphingomyelin (SM) in the intact human low-density lipoprotein (LDL) 1H NMR spectrum at 600 MHz enabled the investigation of LDL surface structure and phospholipid-apoB interactions. We have previously shown that a higher proportion of PC headgroups (25-35% of total PC in LDL) compared to SM were tightly bound to apoB and therefore NMR-invisible [Murphy, H. C., et al. (1997) Biochem. Biophys. Res. Commun. 234 (3), 733-737]. In the present study, we have investigated the mobility of phospholipid (PL) headgroups, using 1H NMR spin-spin (T2) relaxation measurements, in LDL isolated from nine volunteers. We show that both PC and SM exist in two additional and distinct environments indicated by the biexponential behavior of the relaxation decays in each case. The data showed that 36% of PC headgroups had a short T2 component, mean T2 of 31 ms, and 64% had a longer T2 component of 54 ms. Approximately 15% of SM headgroups had a short T2 component (mean T2 of 27 ms) and 85% had a longer T2 component of 78 ms. Therefore the majority of SM headgroups (85%) were more mobile than PC (P < 0.001) and since PC headgroups in organic media were more mobile than SM, we conclude that the characteristic high mobility of LDL SM is not an intrinsic property but arises from a high degree of order in molecular packing of the surface PL of human LDL. We suggest that because PC and SM interact differentially with cholesterol and possibly with neighboring phospholipids, this results in the formation of relatively long-lived microdomains of PL in vivo.
Original languageEnglish
Pages (from-to)9763-9770
Number of pages8
JournalBiochemistry
Volume39
Issue number32
DOIs
Publication statusPublished - 20 Jul 2000
Externally publishedYes

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