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Single-molecule imaging of l-type Ca(2+) channels in live cells
Authors:Harms G S  Cognet L  Lommerse P H  Blab G A  Kahr H  Gamsjäger R  Spaink H P  Soldatov N M  Romanin C  Schmidt T
Affiliation:Department of Biophysics, Leiden University, 2333 CA Leiden, The Netherlands.
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.
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