Clustering of connexin 43-enhanced green fluorescent protein gap junction channels and functional coupling in living cells.
Communication-incompetent cell lines were transfected with connexin (Cx) 43 fused with enhanced green fluorescent protein (EGFP) to examine the relation between Cx distribution determined by fluorescence microscopy and electrical coupling measured at single-channel resolution in living cell pairs. Cx43-EGFP channel properties were like those of wild-type Cx43 except for reduced sensitivity to transjunctional voltage. Cx43-EGFP clustered into plaques at locations of cell-cell contact. Coupling was always absent in the absence of plaques and even in the presence of small plaques. Plaques exceeding several hundred channels always conferred coupling, but only a small fraction of channels were functional. These data indicate that clustering may be a requirement for opening of gap junction channels.
Bukauskas
F F
FF
Department of Neuroscience, Albert Einstein College of Medicine, New York, NY 10461, USA. fbukausk@aecom.yu.edu
Jordan
K
K
Bukauskiene
A
A
Bennett
M V
MV
Lampe
P D
PD
Laird
D W
DW
Verselis
V K
VK
D000818:Animals; D002450:Cell Communication; D002460:Cell Line; D018031:Connexin 43 / Q000378:metabolism*; D004594:Electrophysiology; D017629:Gap Junctions / Q000378:metabolism* / Q000502:physiology; D049452:Green Fluorescent Proteins; D006367:HeLa Cells; D006801:Humans; D008164:Luminescent Proteins / Q000378:metabolism*; D051379:Mice; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D013997:Time Factors; D014162:Transfection; D014407:Tumor Cells, Cultured
D018031:Connexin 43; D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins