CD4 interacts constitutively with multiple CCR5 at the plasma membrane of living cells. A fluorescence recovery after photobleaching at variable radii approach

J Biol Chem. 2007 Nov 30;282(48):35163-8. doi: 10.1074/jbc.M705617200. Epub 2007 Sep 13.

Abstract

The entry of human immunodeficiency virus into target cells requires successive interactions of the viral envelope glycoprotein gp120 with CD4 and the chemokine receptors CCR5 or CXCR4. We previously demonstrated, by Förster resonance energy transfer experiments, the constitutive association of CD4 and CCR5 at the surface of living cells. We therefore speculated that this interaction may correlate with compartmentalization of CD4 and CCR5 within the plasma membrane. Here, we characterize the lateral distribution, the dynamics, and the stoichiometry of these receptors in living cells stably expressing CD4 and/or CCR5 by means of fluorescence recovery after photobleaching at variable radii experiments. We found that (i) these receptors expressed alone are confined into 1-microm-sized domains, (ii) CD4-CCR5 associations occur outside and inside smaller domains, and (iii) these interactions involve multiple CCR5 molecules per CD4.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biophysics / methods
  • CD4 Antigens / biosynthesis*
  • Cell Line
  • Cell Membrane / metabolism*
  • DNA, Complementary / metabolism
  • Diffusion
  • Fluorescence Resonance Energy Transfer / methods
  • Gene Expression Regulation
  • Green Fluorescent Proteins / metabolism
  • Humans
  • Microscopy, Confocal
  • Models, Biological
  • Protein Binding
  • Receptors, CCR5 / metabolism*

Substances

  • CD4 Antigens
  • DNA, Complementary
  • Receptors, CCR5
  • Green Fluorescent Proteins