Title:	Pathomechanismen von HERG-Ionenkanal-Mutationen als Ursache von menschlichen Repolarisationsstoerungen
Authors:	Bertrand, Jessica
Thesis advisor:	Prof. Dr. Eric Schulze-Bahr
Thesis referee:	Prof. Dr. Karl-Heinz Altendorf
Abstract:	Inherited long-QT syndrome is caused by mutations in HERG gene that are associated with distinct mechanisms of ion channel dysfunction (haploinsufficiency or IKr current suppression). Recently, mutations with a gain of HERG channel dysfunction were reported to cause ventricular fibrillation or short-QT syndrome. In the present work, we performed clinical characterization of arrhythmia patients, genotyping and biochemical analysis of HERG mutants in order to elucidate potential disease mechanisms. Using site-directed mutagenesis, 7 identified mutations were inserted into the WT-HERG cDNA. Western blot was used to analyze mutant HERG glycosylation patterns, immunostaining and confocal laser microscopy was performed to localize mutant proteins in different cell compartments. Heterologous expression in Xenopus oocytes was used to analyze IKr currents with the voltage clamp method. The cellular turnover of mutant HERG channels was assessed with pulse-chase experiments. Mutations in the cytoplasmic domains (PAS and cNBD) and in the voltage sensor are trafficking deficient and were identified in LQT2 patients. Three mutations in the N- and C-terminal linker regions undergo regular trafficking to the plasma membrane and were identified compound heterozygous with one of the other mutations in LQT2 patients or separate in patients with IVF. HERG-mutations are associated with various phenotypes like LQT2 and IVF. It seems that there is a direct correlation between the functionality of the protein region with the clinical and molecular biological phenotype. Mutations in functional regions like the PAS- and cNBD-domain lead to a trafficking defect of the mutant proteins and for that reason to a reduction of Ikr. Mutations in less functional regions like the N and C-terminal linker regions undergo normal trafficking and lead to IVF.
URL:	https://osnadocs.ub.uni-osnabrueck.de/handle/urn:nbn:de:gbv:700-2008011414
Subject Keywords:	Ionenkanal; HERG; Mutationen; IKr; Voltage-Clamp; Trafficking
Issue Date:	11-Jan-2008
Submission date:	11-Jan-2008
Type of publication:	Dissertation oder Habilitation [doctoralThesis]
Appears in Collections:	FB05 - E-Dissertationen

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