Intracellular interactions between protein 4.1 and glycophorin C on transport vesicles, as determined by fluorescence correlation spectroscopy

FEBS Lett. 2012 Mar 23;586(6):668-74. doi: 10.1016/j.febslet.2012.01.058. Epub 2012 Feb 9.

Abstract

Interaction of protein 4.1 (4.1R) with the transmembrane protein glycophorin C (GPC) regulates the functions of erythrocyte membrane. Fluorescence correlation spectroscopy (FCS) was used to define the interaction of EGFP-4.1R with DsRed-GPC on transport vesicles (TVs) by measuring their fluctuation in living cells. DsRed-GPC expressed in HeLa cells was delivered to the plasma membrane through slow vesicle transport. EGFP-4.1R, which freely diffused in the cytosol when expressed alone, diffused slowly when co-expressed with DsRed-GPC, indicating association of EGFP-4.1R with TVs. Fluorescence cross-correlation spectroscopy (FCCS) showed direct interaction of EGFP-4.1R with DsRed-GPC on TVs. The present study demonstrates that 4.1R binds to GPC on TVs in living cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Membrane / metabolism
  • Cytoskeletal Proteins / chemistry*
  • Cytoskeletal Proteins / metabolism*
  • Fluorescent Dyes / chemistry
  • Fluorescent Dyes / metabolism
  • Glycophorins / chemistry
  • Glycophorins / genetics
  • Glycophorins / metabolism*
  • Green Fluorescent Proteins / chemistry
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • HeLa Cells
  • Humans
  • Membrane Proteins / chemistry
  • Membrane Proteins / genetics
  • Membrane Proteins / metabolism*
  • Protein Binding
  • Spectrometry, Fluorescence / methods*
  • Transport Vesicles / chemistry*
  • Transport Vesicles / metabolism

Substances

  • Cytoskeletal Proteins
  • Fluorescent Dyes
  • Glycophorins
  • Membrane Proteins
  • enhanced green fluorescent protein
  • erythrocyte membrane band 4.1 protein
  • Green Fluorescent Proteins