Colocalization and nonrandom distribution of Kv1.3 potassium channels and CD3 molecules in the plasma membrane of human T lymphocytes.
Distribution and lateral organization of Kv1.3 potassium channels and CD3 molecules were studied by using electron microscopy, confocal laser scanning microscopy, and fluorescence resonance energy transfer. Immunogold labeling and electron microscopy showed that the distribution of FLAG epitope-tagged Kv1.3 channels (Kv1.3/FLAG) significantly differs from the stochastic Poisson distribution in the plasma membrane of human T lymphoma cells. Confocal laser scanning microscopy images showed that Kv1.3/FLAG channels and CD3 molecules accumulated in largely overlapping membrane areas. The numerical analysis of crosscorrelation of the spatial intensity distributions yielded a high correlation coefficient (C = 0.64). A different hierarchical level of molecular proximity between Kv1.3/FLAG and CD3 proteins was reported by a high fluorescence resonance energy transfer efficiency (E = 51%). These findings implicate that reciprocal regulation of ion-channel activity, membrane potential, and the function of receptor complexes may contribute to the proper functioning of the immunological synapse.
Panyi
G
G
Department of Biophysics and Cell Biology, Research Center for Molecular Medicine, Medical and Health Science Center, University of Debrecen, Nagyerdei krt. 98, H-4012, Debrecen, Hungary.
Bagdány
M
M
Bodnár
A
A
Vámosi
G
G
Szentesi
G
G
Jenei
A
A
Mátyus
L
L
Varga
S
S
Waldmann
T A
TA
Gáspar
R
R
Damjanovich
S
S
D000818:Animals; D017252:CD3 Complex / Q000096:biosynthesis*; D002462:Cell Membrane / Q000276:immunology / Q000378:metabolism*; D004594:Electrophysiology; D000939:Epitopes; D031541:Fluorescence Resonance Energy Transfer; D006801:Humans; D007150:Immunohistochemistry; D019169:Jurkat Cells; D051664:Kv1.3 Potassium Channel; D051379:Mice; D018613:Microscopy, Confocal; D008854:Microscopy, Electron; D015233:Models, Statistical; D015221:Potassium Channels / Q000096:biosynthesis* / Q000737:chemistry*; D024642:Potassium Channels, Voltage-Gated; D013601:T-Lymphocytes / Q000378:metabolism*; D014162:Transfection
D017252:CD3 Complex; D000939:Epitopes; C496973:KCNA3 protein, human; C496974:Kcna3 protein, mouse; D051664:Kv1.3 Potassium Channel; D015221:Potassium Channels; D024642:Potassium Channels, Voltage-Gated