Colocalization and coassembly of two human brain M-type potassium channel subunits that are mutated in epilepsy.
Acetylcholine excites many central and autonomic neurons through inhibition of M-channels, slowly activating, noninactivating voltage-gated potassium channels. We here provide information regarding the in vivo distribution and biochemical characteristics of human brain KCNQ2 and KCNQ3, two channel subunits that form M-channels when expressed in vitro, and, when mutated, cause the dominantly inherited epileptic syndrome, benign neonatal familial convulsions. KCNQ2 and KCNQ3 proteins are colocalized in a somatodendritic pattern on pyramidal and polymorphic neurons in the human cortex and hippocampus. Immunoreactivity for KCNQ2, but not KCNQ3, is also prominent in some terminal fields, suggesting a presynaptic role for a distinct subgroup of M-channels in the regulation of action potential propagation and neurotransmitter release. KCNQ2 and KCNQ3 can be coimmunoprecipitated from brain lysates. Further, KCNQ2 and KCNQ3 are coassociated with tubulin and protein kinase A within a Triton X-100-insoluble protein complex. This complex is not associated with low-density membrane rafts or with N-methyl-d-aspartate receptors, PSD-95 scaffolding proteins, or other potassium channels tested. Our studies thus provide a view of a signaling complex that may be important for cognitive function as well as epilepsy. Analysis of this complex may shed light on the unknown transduction pathway linking muscarinic acetylcholine receptor activation to M-channel inhibition.
Cooper
E C
EC
Department of Neurology, Howard Hughes Medical Institute, University of California, San Francisco, CA 94143, USA.
Aldape
K D
KD
Abosch
A
A
Barbaro
N M
NM
Berger
M S
MS
Peacock
W S
WS
Jan
Y N
YN
Jan
L Y
LY
D001921:Brain / Q000378:metabolism* / Q000473:pathology; D002460:Cell Line; D002540:Cerebral Cortex / Q000378:metabolism / Q000473:pathology; D004827:Epilepsy / Q000235:genetics* / Q000473:pathology; D006624:Hippocampus / Q000378:metabolism / Q000473:pathology; D006801:Humans; D007150:Immunohistochemistry; D051658:KCNQ2 Potassium Channel; D051659:KCNQ3 Potassium Channel; D046911:Macromolecular Substances; D015221:Potassium Channels / Q000032:analysis* / Q000235:genetics*; D024642:Potassium Channels, Voltage-Gated; D011994:Recombinant Proteins / Q000032:analysis / Q000096:biosynthesis; D013347:Subcellular Fractions / Q000378:metabolism; D014162:Transfection
D051658:KCNQ2 Potassium Channel; C496914:KCNQ2 protein, human; D051659:KCNQ3 Potassium Channel; C496916:KCNQ3 protein, human; D046911:Macromolecular Substances; D015221:Potassium Channels; D024642:Potassium Channels, Voltage-Gated; D011994:Recombinant Proteins