Revealing the mechanism of passive transport in lipid bilayers via phonon-mediated nanometre-scale density fluctuations

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Abstract

The passive transport of molecules through a cell membrane relies on thermal motions of the lipids. However, the nature of transmembrane transport and the precise mechanism remain elusive and call for a comprehensive study of phonon excitations. Here we report a high resolution inelastic X-ray scattering study of the in-plane phonon excitations in 1,2-dipalmitoyl-sn-glycero-3-phosphocholine above and below the main transition temperature. In the gel phase, for the first time, we observe low-frequency transverse modes, which exhibit a phonon gap when the lipid transitions into the fluid phase. We argue that the phonon gap signifies the formation of short-lived nanometre-scale lipid clusters and transient pores, which facilitate the passive molecular transport across the bilayer plane. Our findings suggest that the phononic motion of the hydrocarbon tails provides an effective mechanism of passive transport, and illustrate the importance of the collective dynamics of biomembranes.

Figures

  • Figure 1 | Geometry of IXS experiment and DPPC structure factor. (a) In-plane scattering geometry used at the IXS beamline (ID28) at ESRF. The incident and scattered beams remain within the lipid membrane plane and thus the momentum transfer vector Q lies parallel to the sample surface. (b) S(Q,0) as measured showing the chain correlation peak for 45 C. ESRF, European Synchrotron Radiation Facility.
  • Figure 2 | IXS spectra of DPPC. (a) IXS spectra measured at T¼45 C in the fluid phase and at (b) T¼ 20 C in the gel phase. The experimental data (black squares) with error bars signifying 1 s.d. are reported together with the best least squares fit (red solid curves) using a DHO model with two
  • Figure 3 | Propagating transverse excitations. Experimental data, the fitting curve and excitations from Fig. 2 for Q¼ 3.89 nm 1 multiplied by (:o)2 for 45 C (a) and 20 C (b), respectively. In a, the experimental data are well described by the sum of the RF and a single, longitudinal excitation
  • Figure 4 | Statistical analysis of the IXS spectra at Q¼ 3.89 nm 1. (a–d) The best model fits of the IXS scans at 20 C (a,b) and 45 C (c,d) at Q¼ 3.89 nm 1 with one (longitudinal) and two (longitudinal and transverse) DHO excitations, respectively. The residuals are shown by blue dash-dotted curves. The resulting reduced w2 is indicated in each panel. The error bars represent one s.d.
  • Figure 5 | Phonon dispersion curves and damping ratios. (a) o(Q) dispersion curves obtained using the DHO modelling. Filled and open symbols correspond to dispersions curves for 20 and 45 C, respectively. Emergence of high-frequency transverse phonon gap at high temperature is indicated by the arrow. The solid lines are shown to guide eyes only. The dashed lines extrapolate the longitudinal phonon dispersions to Q¼0. (b) The values of the damping ratio R¼o(Q)/G(Q) of the longitudinal and transverse branches are reported as a function of Q, at 20 C. The solid lines through the symbols are guide to eyes. The inset displays R for the 45 C. Since the R values for transverse phonons at 45 C shows no specific trend (scattered near 1) the solid line was omitted. Errors bars represent 1 s.e.m.
  • Figure 6 | Formation of local DPPC clusters and pores. The schematic representation of the lipid bilayer transition from the Lb’ to La phase. In the Lb’ phase the lipids are tightly packed and thus the transverse phonons are supported over large distances. When the temperature is increased beyond Tm and the lipid bilayer undergoes the phase transition to La phase, the molecular arrangement is mostly disordered. However, due to thermal fluctuations, the local short-lived (on the order of a picosecond) lipid clustering with a size of B1.1–1.6 nm (or of the order of several lipids across) is triggered. The lipids,
  • Figure 7 | Assessment of the X-ray beam damage of DPPC. The comparison of the raw IXS scans (T¼45 C, Q¼ 3.89 nm 1) at the beginning and at the end of the measurement with the total duration of the sample’s exposure to the X-ray beam of B34 h. The error bars

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Zhernenkov, M., Bolmatov, D., Soloviov, D., Zhernenkov, K., Toperverg, B. P., Cunsolo, A., … Cai, Y. Q. (2016). Revealing the mechanism of passive transport in lipid bilayers via phonon-mediated nanometre-scale density fluctuations. Nature Communications, 7. https://doi.org/10.1038/ncomms11575

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