Partition of membrane probes in a gel/fluid two-component lipid system: a fluorescence resonance energy transfer study

Biochim Biophys Acta. 2000 Jul 31;1467(1):101-12. doi: 10.1016/s0005-2736(00)00211-x.

Abstract

A non-ideal lipid binary mixture (dilauroylphosphatidylcholine/distearoylphosphatidylcholine), which exhibits gel/fluid phase coexistence for wide temperature and composition ranges, was studied using photophysical techniques, namely fluorescence anisotropy, lifetime and resonance energy transfer (FRET) measurements. The FRET donor, N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)-dilauroylphosphatidylethanol amine, and a short-tailed FRET acceptor, 1,1'-didodecil-3,3,3',3'-tetramethylindocarbocyanine (DiIC12(3)), were shown to prefer the fluid phase by both intrinsic anisotropy, lifetime and FRET measurements, in agreement with published reports. The other studied FRET acceptor, long-tailed probe 1,1'-dioctadecil-3,3,3',3'-tetramethylindocarbocyanine (DiIC18(3)), is usually reported in the literature as partitioning mainly to the gel. While intrinsic lifetime studies indeed indicated preferential partition of DiIC18(3) into a rigidified environment, FRET analysis pointed to an increased donor-acceptor proximity as a consequence of phase separation. These apparently conflicting results were rationalized on the basis of segregation of DiIC18(3) to the gel/fluid interphase. In order to fluid-located donors sense these interphase-located acceptors, fluid domains should be small (not exceed approximately 10-15 nm). It is concluded that membrane probes which apparently prefer the gel phase may indeed show a non-random distribution in this medium, and tend to locate in an environment which simultaneously leads to less strict packing constraints and to favorable hydrophobic matching interactions.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 4-Chloro-7-nitrobenzofurazan / analogs & derivatives*
  • Carbocyanines
  • Fluorescent Dyes / chemistry*
  • Phosphatidylcholines / chemistry*
  • Phosphatidylethanolamines
  • Spectrometry, Fluorescence
  • Temperature

Substances

  • Carbocyanines
  • Fluorescent Dyes
  • Phosphatidylcholines
  • Phosphatidylethanolamines
  • 1,2-dilauroylphosphatidylcholine
  • 3,3'-dioctadecylindocarbocyanine
  • carbocyanine dye DiIC12(3)
  • N-NBD-1,2-dilauroylphosphatidylethanolamine
  • 1,2-distearoyllecithin
  • 4-Chloro-7-nitrobenzofurazan