title: Trafficking-deficient long QT syndrome mutation KCNQ1-T587M confers severe clinical phenotype by impairment of KCNH2 membrane localization: evidence for clinically significant IKr-IKs alpha-subunit interaction.
abstract: BACKGROUND
  KCNQ1-T587M is a trafficking-deficient long QT syndrome (LQTS) missense mutation. Affected patients exhibit severe clinical phenotypes that are not explained by the mutant's effects on I(Ks). Previous work showed a KCNH2 and KCNQ1 alpha-subunit interaction that increases KCNH2 membrane localization and function.
  
  
  OBJECTIVE
  We hypothesized that failure of trafficking-deficient KCNQ1-T587M to enhance KCNH2 membrane expression could reduce KCNH2 current versus wild-type KCNQ1 (KCNQ1-WT), contributing to the LQTS phenotype of KCNQ1-T587M carriers.
  
  
  METHODS
  Patch-clamp, protein biochemical studies, confocal imaging, and in vivo transfection of guinea pig cardiomyocytes were performed.
  
  
  RESULTS
  KCNQ1-T587M failed to generate functional current when coexpressed with KCNE1 and caused haploinsufficiency when coexpressed with KCNQ1-WT/KCNE1. Coexpression of KCNQ1-WT with KCNH2 increased I(KCNH2) versus KCNH2 alone (P <.05). Immunoblots and confocal microscopy indicated increased plasma membrane localization of KCNH2 alpha-subunits in cells cotransfected with KCNQ1-WT plasmid, while total KCNH2 protein synthesis and KCNH2 glycosylation remained unaffected, which suggests a chaperone effect of KCNQ1-WT to enhance the membrane localization of KCNH2. KCNH2 also coimmunoprecipitated with KCNQ1-WT. Although KCNQ1-T587M coprecipitated with KCNH2, the mutant was retained intracellularly and failed to increase KCNH2 membrane localization, abolishing the KCNQ1-WT chaperone function and reducing I(KCNH2) upon coexpression substantially compared with coexpression with KCNQ1-WT (P <.05). In vivo transfection of KCNQ1-T587M in guinea pigs suppressed I(Kr) in isolated cardiomyocytes.
  
  
  CONCLUSIONS
  The trafficking-deficient LQTS mutation KCNQ1-T587M fails to show the chaperoning function that enhances KCNH2 membrane localization with KCNQ1-WT. This novel mechanism results in reduced I(KCNH2), which would be expected to decrease repolarization reserve and synergize with reduced I(KCNQ1) caused directly by the mutation, potentially explaining the malignant clinical phenotype in affected patients.
authors: Div. of Cardiology, Section of Electrophysiology, Goethe-Universität, Frankfurt, Germany.
authors: Peter
authors: P
authors: Biliczki
authors: Zenawit
authors: Z
authors: Girmatsion
authors: Ralf P
authors: RP
authors: Brandes
authors: Sabine
authors: S
authors: Harenkamp
authors: Bruno
authors: B
authors: Pitard
authors: Flavien
authors: F
authors: Charpentier
authors: Terence E
authors: TE
authors: Hébert
authors: Stefan H
authors: SH
authors: Hohnloser
authors: Isabelle
authors: I
authors: Baró
authors: Stanley
authors: S
authors: Nattel
authors: Joachim R
authors: JR
authors: Ehrlich
mesh_terms: D000704:Analysis of Variance; D000818:Animals; D016466:CHO Cells; D002170:Canada; D002460:Cell Line; D006224:Cricetinae; D003412:Cricetulus; D016757:Death, Sudden, Cardiac / Q000473:pathology; D051638:Ether-A-Go-Go Potassium Channels / Q000235:genetics*; D006168:Guinea Pigs; D006801:Humans; D051657:KCNQ1 Potassium Channel / Q000235:genetics*; D008133:Long QT Syndrome / Q000235:genetics*; D018613:Microscopy, Confocal; D020125:Mutation, Missense; D032383:Myocytes, Cardiac / Q000502:physiology; D010641:Phenotype; D016171:Torsades de Pointes / Q000235:genetics; D014162:Transfection
chemical_list: D051638:Ether-A-Go-Go Potassium Channels; D051657:KCNQ1 Potassium Channel