Biophys J, September 2002, p. 1465-1478, Vol. 83, No. 3

A Solid-State NMR Study of Phospholipid-Cholesterol Interactions: Sphingomyelin-Cholesterol Binary Systems

Wen Guo,^* Volker Kurze, Thomas Huber, Nezam H. Afdhal,^§ Klaus Beyer,^¶ and James A. Hamilton

Departments of  ^*Medicine and  Biophysics, Boston University School of Medicine, Boston, Massachusetts 02118 USA;  Department of Radiology, Technische Universität München, Munich, Germany;  Department of Chemistry, University of Arizona, Tucson, Arizona 85721, USA;  ^§Beth Israel Hospital, Harvard Medical School, Boston, Massachusetts 02115, USA; and  ^¶Lehrstuhl für Stoffwechselbiochemie, Ludwig-Maximilians-Universität München, Munich, Germany

We used solid-state NMR techniques to probe the interactions of cholesterol (Chol) with bovine brain sphingomyelin (SM) and for comparison of the interactions of Chol with dipalmitoylphosphatidylcholine (DPPC), which has a similar gel-to-liquid crystalline transition temperature. ¹H-, ³¹P-, and ¹³C-MASNMR yielded high-resolution spectra from multilamellar dispersions of unlabeled brain SM and Chol for analysis of chemical shifts and linewidths. In addition, ²H-NMR spectra of oriented lipid membranes with specific deuterium labels gave information about membrane ordering and mobility. Chol disrupted the gel-phase of pure SM and increased acyl chain ordering in the liquid crystalline phase. As inferred from ¹³C chemical shifts, the boundaries between the ordered and disordered liquid crystalline phases (L and L) were similar for SM and DPPC. The solubility limit of Chol in SM was ~50 mol %, the same value as previously reported for DPPC membranes. We found no evidence for specific H-bonding between Chol and the amide group of SM. The order parameters of a probe molecule, d31-sn1-DPPC, in SM were slightly higher than in DPPC for all carbons except the terminal groups at 30 mol % but were not significantly different at 5 and 60 mol % Chol. These studies show a general similarity with some subtle differences in the way Chol interacts with DPPC and SM. In the environment of a typical biomembrane, the higher proportion of saturated fatty acyl chains in SM compared to other phospholipids may be the most significant factor influencing interactions with Chol.

Biophys J, September 2002, p. 1465-1478, Vol. 83, No. 3
© 2002 by the Biophysical Society   0006-3495/02/09/1465/14  $2.00

This article has been cited by other articles:

				G. P. Holland and T. M. Alam Unique Backbone-Water Interaction Detected in Sphingomyelin Bilayers with 1H/31P and 1H/13C HETCOR MAS NMR Spectroscopy Biophys. J., August 1, 2008; 95(3): 1189 - 1198. [Abstract] [Full Text] [PDF]

				S. P. Soni, D. S. LoCascio, Y. Liu, J. A. Williams, R. Bittman, W. Stillwell, and S. R. Wassall Docosahexaenoic Acid Enhances Segregation of Lipids between Raft and Nonraft Domains: 2H-NMR Study Biophys. J., July 1, 2008; 95(1): 203 - 214. [Abstract] [Full Text] [PDF]

				B. M. Stott, M. P. Vu, C. O. McLemore, M. S. Lund, E. Gibbons, T. J. Brueseke, H. A. Wilson-Ashworth, and J. D. Bell Use of fluorescence to determine the effects of cholesterol on lipid behavior in sphingomyelin liposomes and erythrocyte membranes J. Lipid Res., June 1, 2008; 49(6): 1202 - 1215. [Abstract] [Full Text] [PDF]

				J. Aittoniemi, P. S. Niemela, M. T. Hyvonen, M. Karttunen, and I. Vattulainen Insight into the Putative Specific Interactions between Cholesterol, Sphingomyelin, and Palmitoyl-Oleoyl Phosphatidylcholine Biophys. J., February 15, 2007; 92(4): 1125 - 1137. [Abstract] [Full Text] [PDF]

				A. Pokorny, L. E. Yandek, A. I. Elegbede, A. Hinderliter, and P. F. F. Almeida Temperature and Composition Dependence of the Interaction of {delta}-Lysin with Ternary Mixtures of Sphingomyelin/Cholesterol/POPC Biophys. J., September 15, 2006; 91(6): 2184 - 2197. [Abstract] [Full Text] [PDF]

				A. Tsamaloukas, H. Szadkowska, and H. Heerklotz Thermodynamic Comparison of the Interactions of Cholesterol with Unsaturated Phospholipid and Sphingomyelins Biophys. J., June 15, 2006; 90(12): 4479 - 4487. [Abstract] [Full Text] [PDF]

				C. Chachaty, D. Rainteau, C. Tessier, P. J. Quinn, and C. Wolf Building Up of the Liquid-Ordered Phase Formed by Sphingomyelin and Cholesterol Biophys. J., June 1, 2005; 88(6): 4032 - 4044. [Abstract] [Full Text] [PDF]

				B. L. Stottrup, D. S. Stevens, and S. L. Keller Miscibility of Ternary Mixtures of Phospholipids and Cholesterol in Monolayers, and Application to Bilayer Systems Biophys. J., January 1, 2005; 88(1): 269 - 276. [Abstract] [Full Text] [PDF]

				P. Niemela, M. T. Hyvonen, and I. Vattulainen Structure and Dynamics of Sphingomyelin Bilayer: Insight Gained through Systematic Comparison to Phosphatidylcholine Biophys. J., November 1, 2004; 87(5): 2976 - 2989. [Abstract] [Full Text] [PDF]

				S. A. Pandit, S. Vasudevan, S. W. Chiu, R. J. Mashl, E. Jakobsson, and H. L. Scott Sphingomyelin-Cholesterol Domains in Phospholipid Membranes: Atomistic Simulation Biophys. J., August 1, 2004; 87(2): 1092 - 1100. [Abstract] [Full Text] [PDF]

				J. M. Holopainen, A. J. Metso, J.-P. Mattila, A. Jutila, and P. K. J. Kinnunen Evidence for the Lack of a Specific Interaction between Cholesterol and Sphingomyelin Biophys. J., March 1, 2004; 86(3): 1510 - 1520. [Abstract] [Full Text] [PDF]

				M. Koivusalo, J. Alvesalo, J. A. Virtanen, and P. Somerharju Partitioning of Pyrene-Labeled Phospho- and Sphingolipids between Ordered and Disordered Bilayer Domains Biophys. J., February 1, 2004; 86(2): 923 - 935. [Abstract] [Full Text] [PDF]

				R. F. M. de Almeida, A. Fedorov, and M. Prieto Sphingomyelin/Phosphatidylcholine/Cholesterol Phase Diagram: Boundaries and Composition of Lipid Rafts Biophys. J., October 1, 2003; 85(4): 2406 - 2416. [Abstract] [Full Text] [PDF]

				A. Filippov, G. Oradd, and G. Lindblom The Effect of Cholesterol on the Lateral Diffusion of Phospholipids in Oriented Bilayers Biophys. J., May 1, 2003; 84(5): 3079 - 3086. [Abstract] [Full Text] [PDF]

				R. M. Epand Cholesterol in Bilayers of Sphingomyelin or Dihydrosphingomyelin at Concentrations Found in Ocular Lens Membranes Biophys. J., May 1, 2003; 84(5): 3102 - 3110. [Abstract] [Full Text] [PDF]