N-Glycosylation of TREK-1/hK2P2.1 Two-Pore-Domain Potassium (K2P) Channels
Mechanosensitive hTREK-1 two-pore-domain potassium (hK2P2.1) channels give rise to background currents that control cellular excitability. Recently, TREK-1 currents have been linked to the regulation of cardiac rhythm as well as to hypertrophy and fibrosis. Even though the pharmacological and biophysical characteristics of hTREK-1 channels have been widely studied, relatively little is known about their posttranslational modifications. This study aimed to evaluate whether hTREK-1 channels are N-glycosylated and whether glycosylation may affect channel functionality. Following pharmacological inhibition of N-glycosylation, enzymatic digestion or mutagenesis, immunoblots of Xenopus laevis oocytes and HEK-293T cell lysates were used to assess electrophoretic mobility. Two-electrode voltage clamp measurements were employed to study channel function. TREK-1 channel subunits undergo N-glycosylation at asparagine residues 110 and 134. The presence of sugar moieties at these two sites increases channel function. Detection of glycosylation-deficient mutant channels in surface fractions and recordings of macroscopic potassium currents mediated by these subunits demonstrated that nonglycosylated hTREK-1 channel subunits are able to reach the cell surface in general but with seemingly reduced efficiency compared to glycosylated subunits. These findings extend our understanding of the regulation of hTREK-1 currents by posttranslational modifications and provide novel insights into how altered ion channel glycosylation may promote arrhythmogenesis.
| Medienart: |
E-Artikel |
|---|
| Erscheinungsjahr: |
2019 |
|---|---|
| Erschienen: |
2019 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:20 |
|---|---|
| Enthalten in: |
International journal of molecular sciences - 20(2019), 20 vom: 20. Okt. |
| Sprache: |
Englisch |
|---|
| Beteiligte Personen: |
Wiedmann, Felix [VerfasserIn] |
|---|
| Links: |
|---|
| Anmerkungen: |
Date Completed 25.03.2020 Date Revised 23.06.2020 published: Electronic Citation Status MEDLINE |
|---|
| doi: |
10.3390/ijms20205193 |
|---|
| funding: |
|
|---|---|
| Förderinstitution / Projekttitel: |
|
| PPN (Katalog-ID): |
NLM302413782 |
|---|
| LEADER | 01000naa a22002652 4500 | ||
|---|---|---|---|
| 001 | NLM302413782 | ||
| 003 | DE-627 | ||
| 005 | 20231225110656.0 | ||
| 007 | cr uuu---uuuuu | ||
| 008 | 231225s2019 xx |||||o 00| ||eng c | ||
| 024 | 7 | |a 10.3390/ijms20205193 |2 doi | |
| 028 | 5 | 2 | |a pubmed24n1008.xml |
| 035 | |a (DE-627)NLM302413782 | ||
| 035 | |a (NLM)31635148 | ||
| 035 | |a (PII)E5193 | ||
| 040 | |a DE-627 |b ger |c DE-627 |e rakwb | ||
| 041 | |a eng | ||
| 100 | 1 | |a Wiedmann, Felix |e verfasserin |4 aut | |
| 245 | 1 | 0 | |a N-Glycosylation of TREK-1/hK2P2.1 Two-Pore-Domain Potassium (K2P) Channels |
| 264 | 1 | |c 2019 | |
| 336 | |a Text |b txt |2 rdacontent | ||
| 337 | |a ƒaComputermedien |b c |2 rdamedia | ||
| 338 | |a ƒa Online-Ressource |b cr |2 rdacarrier | ||
| 500 | |a Date Completed 25.03.2020 | ||
| 500 | |a Date Revised 23.06.2020 | ||
| 500 | |a published: Electronic | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a Mechanosensitive hTREK-1 two-pore-domain potassium (hK2P2.1) channels give rise to background currents that control cellular excitability. Recently, TREK-1 currents have been linked to the regulation of cardiac rhythm as well as to hypertrophy and fibrosis. Even though the pharmacological and biophysical characteristics of hTREK-1 channels have been widely studied, relatively little is known about their posttranslational modifications. This study aimed to evaluate whether hTREK-1 channels are N-glycosylated and whether glycosylation may affect channel functionality. Following pharmacological inhibition of N-glycosylation, enzymatic digestion or mutagenesis, immunoblots of Xenopus laevis oocytes and HEK-293T cell lysates were used to assess electrophoretic mobility. Two-electrode voltage clamp measurements were employed to study channel function. TREK-1 channel subunits undergo N-glycosylation at asparagine residues 110 and 134. The presence of sugar moieties at these two sites increases channel function. Detection of glycosylation-deficient mutant channels in surface fractions and recordings of macroscopic potassium currents mediated by these subunits demonstrated that nonglycosylated hTREK-1 channel subunits are able to reach the cell surface in general but with seemingly reduced efficiency compared to glycosylated subunits. These findings extend our understanding of the regulation of hTREK-1 currents by posttranslational modifications and provide novel insights into how altered ion channel glycosylation may promote arrhythmogenesis | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a K2P2.1 | |
| 650 | 4 | |a KCNK2 | |
| 650 | 4 | |a N-glycosylation | |
| 650 | 4 | |a TREK-1 | |
| 650 | 4 | |a ion channel | |
| 650 | 4 | |a membrane trafficking | |
| 650 | 4 | |a two-pore-domain potassium channels | |
| 650 | 7 | |a Potassium Channels, Tandem Pore Domain |2 NLM | |
| 650 | 7 | |a potassium channel protein TREK-1 |2 NLM | |
| 650 | 7 | |a Potassium |2 NLM | |
| 650 | 7 | |a RWP5GA015D |2 NLM | |
| 700 | 1 | |a Schlund, Daniel |e verfasserin |4 aut | |
| 700 | 1 | |a Faustino, Francisco |e verfasserin |4 aut | |
| 700 | 1 | |a Kraft, Manuel |e verfasserin |4 aut | |
| 700 | 1 | |a Ratte, Antonius |e verfasserin |4 aut | |
| 700 | 1 | |a Thomas, Dierk |e verfasserin |4 aut | |
| 700 | 1 | |a Katus, Hugo A |e verfasserin |4 aut | |
| 700 | 1 | |a Schmidt, Constanze |e verfasserin |4 aut | |
| 773 | 0 | 8 | |i Enthalten in |t International journal of molecular sciences |d 2008 |g 20(2019), 20 vom: 20. Okt. |w (DE-627)NLM185552161 |x 1422-0067 |7 nnns |
| 773 | 1 | 8 | |g volume:20 |g year:2019 |g number:20 |g day:20 |g month:10 |
| 856 | 4 | 0 | |u http://dx.doi.org/10.3390/ijms20205193 |3 Volltext |
| 912 | |a GBV_USEFLAG_A | ||
| 912 | |a GBV_NLM | ||
| 951 | |a AR | ||
| 952 | |d 20 |j 2019 |e 20 |b 20 |c 10 | ||