Long QT2 mutation on the Kv11.1 ion channel inhibits current activity by ablating a protein kinase Cα consensus site.
Mutations that inhibit Kv11.1 ion channel activity contribute to abnormalities of cardiac repolarization that can lead to long QT2 (LQT2) cardiac arrhythmias and sudden death. However, for most of these mutations, nothing is known about the molecular mechanism linking Kv11.1 malfunction to cardiac death. We have previously demonstrated that disease-related mutations that create consensus sites for kinases on ion channels can dramatically change ion channel activity. Here, we show that a LQT2-associated mutation can inhibit Kv11.1 ion channel activity by perturbing a consensus site for the Ser/Thr protein kinase C α (PKCα). We first reveal by mass spectrometry analysis that Ser890 of the Kv11.1 ion channel is phosphorylated. Then, we demonstrate by a phospho-detection immunoassay combined with genetic manipulation that PKCα phosphorylates Ser890. Furthermore, we show that Ser890 phosphorylation is associated with an increase in Kv11.1 membrane density with alteration of recovery from inactivation. In addition, a newly discovered and as yet uncharacterized LQT2-associated nonsynonymous single nucleotide polymorphism 2660 G→A within the human ether-á-go-go-related gene 1 coding sequence, which replaces arginine 887 with a histidine residue (R887H), strongly inhibits PKCα-dependent phosphorylation of residue Ser890 on Kv11.1, and ultimately inhibits surface expression and current density. Taken together, our data provide a functional link between this channel mutation and LQT2.
Departments of Molecular Pharmacology and Therapeutics, Loyola University, Chicago, Illinois, USA.
Alexander J
AJ
Donovan
Katherine
K
Lansu
Jason G
JG
Williams
Mitchell F
MF
Denning
Saverio
S
Gentile
D000818:Animals; D016466:CHO Cells; D002478:Cells, Cultured; D016384:Consensus Sequence; D006224:Cricetinae; D000072237:ERG1 Potassium Channel; D051638:Ether-A-Go-Go Potassium Channels / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D007473:Ion Channels / Q000235:genetics* / Q000378:metabolism; D008133:Long QT Syndrome / Q000235:genetics* / Q000378:metabolism; D009154:Mutation; D010766:Phosphorylation / Q000235:genetics; D051571:Protein Kinase C-alpha / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism; D059748:Proteolysis; D013329:Structure-Activity Relationship
D000072237:ERG1 Potassium Channel; D051638:Ether-A-Go-Go Potassium Channels; D007473:Ion Channels; C000606913:KCNH2 protein, human; D051571:Protein Kinase C-alpha