Apoptotic surface delivery of K+ channels.
Apoptosis in cortical neurons requires efflux of cytoplasmic potassium mediated by a surge in Kv2.1 channel activity. Pharmacological blockade or molecular disruption of these channels in neurons prevents apoptotic cell death, while ectopic expression of Kv2.1 channels promotes apoptosis in non-neuronal cells. Here, we use a cysteine-containing mutant of Kv2.1 and a thiol-reactive covalent inhibitor to demonstrate that the increase in K+ current during apoptosis is due to de novo insertion of functional channels into the plasma membrane. Biotinylation experiments confirmed the delivery of additional Kv2.1 protein to the cell surface following an apoptotic stimulus. Finally, expression of botulinum neurotoxins that cleave syntaxin and synaptosome-associated protein of 25 kDa (SNAP-25) blocked upregulation of surface Kv2.1 channels in cortical neurons, suggesting that target soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) proteins support proapoptotic delivery of K+ channels. These data indicate that trafficking of Kv2.1 channels to the plasma membrane causes the apoptotic surge in K+ current.
Pal
S K
SK
Department of Neurobiology, University of Pittsburgh School of Medicine, Pittsburgh, PA 15217, USA.
Takimoto
K
K
Aizenman
E
E
Levitan
E S
ES
D000818:Animals; D017209:Apoptosis; D019921:Biotinylation; D016466:CHO Cells; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism*; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000196:embryology; D006224:Cricetinae; D003412:Cricetulus; D008564:Membrane Potentials; D009474:Neurons / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D011188:Potassium / Q000378:metabolism; D011725:Pyridines / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley; D050600:SNARE Proteins / Q000378:metabolism; D051669:Shab Potassium Channels / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D014162:Transfection
D011725:Pyridines; D050600:SNARE Proteins; D051669:Shab Potassium Channels; D011188:Potassium