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Two-pore-domain potassium (K) channels mediate potassium background currents that stabilize the resting membrane potential and facilitate action potential repolarization. In the human heart, hK17.1 channels are predominantly expressed in the atria and Purkinje cell. Reduced atrial hK17.1 protein levels were described in patients with atrial fibrillation or heart failure. Genetic alterations in hK17.1 were associated with cardiac conduction disorders. Little is known about posttranslational modifications of hK17.1. Here, we characterized glycosylation of hK17.1 and investigated how glycosylation alters its surface expression and activity. Wild-type (WT) hK17.1 channels and channels lacking specific glycosylation sites, were expressed in Xenopus laevis oocytes, HEK-293T cells and HeLa cells. N-glycosylation was disrupted using N-glycosidase F and tunicamycin. hK17.1 expression and activity were assessed using immunoblot analysis and two-electrode voltage-clamp technique. hK17.1 channel subunits harbor two functional N-glycosylation sites at position N65 and N94. In hemi-glycosylated hK17.1 channels, functionality and membrane trafficking remain preserved. Disruption of both N-glycosylation sites results in loss of hK17.1 currents, presumably caused by impaired surface expression. This study confirms di-glycosylation of hK17.1 channel subunits and its pivotal role in cell surface targeting. Our findings underline the functional relevance of N-glycosylation in biogenesis and membrane trafficking of ion channels.
This article was published in the following journal.
Name: Molecular biology of the cell
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ATP-dependent DNA helicase that contains two N-terminal ZINC FINGERS and C-terminal ATP-binding and helicase domains. It functions in the regulation of gene transcription and CHROMATIN REMODELING. ATRX undergoes cell-cycle dependent phosphorylation, which causes it to translocate from the NUCLEAR MATRIX to CHROMATIN; thus, it may change its role from gene regulation during INTERPHASE to ensuring proper chromosome segregation at MITOSIS. Mutations in the ATRX gene are associated with cases of X-LINKED MENTAL RETARDATION co-morbid with ALPHA-THALASSEMIA (ATRX syndrome).
A CYCLIN C dependent kinase that is an important component of the mediator complex. The enzyme is activated by its interaction with CYCLIN C and plays a role in transcriptional regulation by phosphorylating RNA POLYMERASE II.
A cyclin-dependent kinase that forms a complex with CYCLIN C and is active during the G1 PHASE of the CELL CYCLE. It plays a role in the transition from G1 to S PHASE and in transcriptional regulation.
Cardiology is a specialty of internal medicine. Cardiac electrophysiology : Study of the electrical properties and conduction diseases of the heart. Echocardiography : The use of ultrasound to study the mechanical function/physics of the h...