Single-molecule imaging of l-type Ca(2+) channels in live cells

Biophys J. 2001 Nov;81(5):2639-46. doi: 10.1016/S0006-3495(01)75907-3.

Abstract

L-type Ca(2+) channels are an important means by which a cell regulates the Ca(2+) influx into the cytosol on electrical stimulation. Their structure and dynamics in the plasma membrane, including their molecular mobility and aggregation, is of key interest for the in-depth understanding of their function. Construction of a fluorescent variant by fusion of the yellow-fluorescent protein to the ion channel and expression in a human cell line allowed us to address its dynamic embedding in the membrane at the level of individual channels in vivo. We report on the observation of individual fluorescence-labeled human cardiac L-type Ca(2+) channels using wide-field fluorescence microscopy in living cells. Our fluorescence and electrophysiological data indicate that L-type Ca(2+) channels tend to form larger aggregates which are mobile in the plasma membrane.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Proteins / chemistry*
  • Bacterial Proteins / genetics
  • Calcium Channels, L-Type / analysis*
  • Calcium Channels, L-Type / genetics
  • Calcium Channels, L-Type / metabolism
  • Cell Line / cytology
  • Cell Membrane / metabolism
  • Diagnostic Imaging / methods*
  • Electrophysiology / methods
  • Humans
  • Kidney / cytology
  • Luminescent Proteins / chemistry*
  • Luminescent Proteins / genetics
  • Microscopy, Fluorescence / methods
  • Movement / physiology
  • Myocardium / cytology
  • Protein Binding / physiology
  • Recombinant Fusion Proteins / analysis*

Substances

  • Bacterial Proteins
  • Calcium Channels, L-Type
  • Luminescent Proteins
  • Recombinant Fusion Proteins
  • yellow fluorescent protein, Bacteria