title: Single-molecule imaging of l-type Ca(2+) channels in live cells.
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.
authors.affiliation: Department of Biophysics, Leiden University, 2333 CA Leiden, The Netherlands.
authors.forename: G S
authors.initials: GS
authors.lastname: Harms
authors.forename: L
authors.initials: L
authors.lastname: Cognet
authors.forename: P H
authors.initials: PH
authors.lastname: Lommerse
authors.forename: G A
authors.initials: GA
authors.lastname: Blab
authors.forename: H
authors.initials: H
authors.lastname: Kahr
authors.forename: R
authors.initials: R
authors.lastname: Gamsjäger
authors.forename: H P
authors.initials: HP
authors.lastname: Spaink
authors.forename: N M
authors.initials: NM
authors.lastname: Soldatov
authors.forename: C
authors.initials: C
authors.lastname: Romanin
authors.forename: T
authors.initials: T
authors.lastname: Schmidt
mesh_terms: 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*
chemical_list: D001426:Bacterial Proteins; D020746:Calcium Channels, L-Type; D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; C054562:yellow fluorescent protein, Bacteria