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Progressive ordering with decreasing temperature of the phospholipids of influenza virus

Abstract

Using linewidth and spinning sideband intensities of lipid hydrocarbon chain resonances in proton magic angle spinning NMR spectra, we detected the temperature-dependent phase state of naturally occurring lipids of intact influenza virus without exogenous probes. Increasingly, below 41 °C ordered and disordered lipid domains coexisted for the viral envelope and extracts thereof. At 22 °C much lipid was in a gel phase, the fraction of which reversibly increased with cholesterol depletion. Diffusion measurements and fluorescence microscopy independently confirmed the existence of gel-phase domains. Thus the existence of ordered regions of lipids in biological membranes is now demonstrated. Above the physiological temperatures of influenza infection, the physical properties of viral envelope lipids, regardless of protein content, were indistinguishable from those of the disordered fraction. Viral fusion appears to be uncorrelated to ordered lipid content. Lipid ordering may contribute to viral stability at lower temperatures, which has recently been found to be critical for airborne transmission.

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Figure 1: 1H MAS NMR spectra of model and biomembranes.
Figure 2: 1H MAS NMR spectra as a function of temperature.
Figure 3: Fraction of ordered lipids as a function of temperature.
Figure 4: 1H MAS NMR diffusion measurements.
Figure 5: Fluorescence microscopy detection of ordered lipid domains.
Figure 6: Effect of temperature on lipid mixing assay of influenza virus fusion with RBC membranes.

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Acknowledgements

This work was supported by funding from the intramural research programs of the National Institute of Child Health and Human Development and the National Institute on Alcohol Abuse and Alcoholism, US National Institutes of Health. We thank L. Chernomordik and E. Leikina for conducting the experiments on content mixing.

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I.V.P., K.G. and J.Z. designed research, analyzed data and wrote the paper. I.V.P. and L.B. performed research.

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Correspondence to Joshua Zimmerberg.

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Polozov, I., Bezrukov, L., Gawrisch, K. et al. Progressive ordering with decreasing temperature of the phospholipids of influenza virus. Nat Chem Biol 4, 248–255 (2008). https://doi.org/10.1038/nchembio.77

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