Single-molecule imaging of l-type Ca(2+) channels in live cells.
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.
Harms
G S
GS
Department of Biophysics, Leiden University, 2333 CA Leiden, The Netherlands.
Cognet
L
L
Lommerse
P H
PH
Blab
G A
GA
Kahr
H
H
Gamsjäger
R
R
Spaink
H P
HP
Soldatov
N M
NM
Romanin
C
C
Schmidt
T
T
D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics; D020746:Calcium Channels, L-Type / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D002460:Cell Line / Q000166:cytology; D002462:Cell Membrane / Q000378:metabolism; D003952:Diagnostic Imaging / Q000379:methods*; D004594:Electrophysiology / Q000379:methods; D006801:Humans; D007668:Kidney / Q000166:cytology; D008164:Luminescent Proteins / Q000737:chemistry* / Q000235:genetics; D008856:Microscopy, Fluorescence / Q000379:methods; D009068:Movement / Q000502:physiology; D009206:Myocardium / Q000166:cytology; D011485:Protein Binding / Q000502:physiology; D011993:Recombinant Fusion Proteins / Q000032:analysis*
D001426:Bacterial Proteins; D020746:Calcium Channels, L-Type; D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; C054562:yellow fluorescent protein, Bacteria