Transbilayer lipid interactions mediate nanoclustering of lipid-anchored proteins

Cell. 2015 Apr 23;161(3):581-594. doi: 10.1016/j.cell.2015.03.048.


Understanding how functional lipid domains in live cell membranes are generated has posed a challenge. Here, we show that transbilayer interactions are necessary for the generation of cholesterol-dependent nanoclusters of GPI-anchored proteins mediated by membrane-adjacent dynamic actin filaments. We find that long saturated acyl-chains are required for forming GPI-anchor nanoclusters. Simultaneously, at the inner leaflet, long acyl-chain-containing phosphatidylserine (PS) is necessary for transbilayer coupling. All-atom molecular dynamics simulations of asymmetric multicomponent-membrane bilayers in a mixed phase provide evidence that immobilization of long saturated acyl-chain lipids at either leaflet stabilizes cholesterol-dependent transbilayer interactions forming local domains with characteristics similar to a liquid-ordered (lo) phase. This is verified by experiments wherein immobilization of long acyl-chain lipids at one leaflet effects transbilayer interactions of corresponding lipids at the opposite leaflet. This suggests a general mechanism for the generation and stabilization of nanoscale cholesterol-dependent and actin-mediated lipid clusters in live cell membranes.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Actins / metabolism
  • Animals
  • CHO Cells
  • Cell Membrane / metabolism
  • Cricetulus
  • Glycosylphosphatidylinositols / metabolism
  • Lipid-Linked Proteins / metabolism*
  • Molecular Dynamics Simulation
  • Phosphatidylserines / metabolism


  • Actins
  • Glycosylphosphatidylinositols
  • Lipid-Linked Proteins
  • Phosphatidylserines