Gap19, a Cx43 Hemichannel Inhibitor, Acts as a Gating Modifier That Decreases Main State Opening While Increasing Substate Gating
Cx43 hemichannels (HCs) are electrically and chemically gated transmembrane pores with low open probability and multiple conductance states, which makes kinetic studies of channel gating in large datasets challenging. Here, we developed open access software, named HemiGUI, to analyze HC gating trans...
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2020-10-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/21/19/7340 |
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doaj-41c79c6488254f5f94eb4df4e229f7062020-11-25T03:26:58ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672020-10-01217340734010.3390/ijms21197340Gap19, a Cx43 Hemichannel Inhibitor, Acts as a Gating Modifier That Decreases Main State Opening While Increasing Substate GatingAlessio Lissoni0Nan Wang1Timur Nezlobinskii2Maarten De Smet3Alexander V. Panfilov4Nele Vandersickel5Luc Leybaert6Katja Witschas7Department of Basic and Applied Medical Sciences—Physiology Group, Ghent University, 9000 Ghent, BelgiumDepartment of Basic and Applied Medical Sciences—Physiology Group, Ghent University, 9000 Ghent, BelgiumDepartment of Physics and Astronomy, Ghent University, 9000 Ghent, BelgiumDepartment of Basic and Applied Medical Sciences—Physiology Group, Ghent University, 9000 Ghent, BelgiumDepartment of Physics and Astronomy, Ghent University, 9000 Ghent, BelgiumDepartment of Physics and Astronomy, Ghent University, 9000 Ghent, BelgiumDepartment of Basic and Applied Medical Sciences—Physiology Group, Ghent University, 9000 Ghent, BelgiumDepartment of Basic and Applied Medical Sciences—Physiology Group, Ghent University, 9000 Ghent, BelgiumCx43 hemichannels (HCs) are electrically and chemically gated transmembrane pores with low open probability and multiple conductance states, which makes kinetic studies of channel gating in large datasets challenging. Here, we developed open access software, named HemiGUI, to analyze HC gating transitions and investigated voltage-induced HC opening based on up to ≈4000 events recorded in HeLa-Cx43-overexpressing cells. We performed a detailed characterization of Cx43 HC gating profiles and specifically focused on the role of the C-terminal tail (CT) domain by recording the impact of adding an EGFP tag to the Cx43 CT end (Cx43-EGFP) or by supplying the Cx43 HC-inhibiting peptide Gap19 that interferes with CT interaction with the cytoplasmic loop (CL). We found that Gap19 not only decreased HC opening activity to the open state (≈217 pS) but also increased the propensity of subconductance (≈80 pS) transitions that additionally became slower as compared to the control. The work demonstrates that large sample transition analysis allows detailed investigations on Cx43 HC gating and shows that Gap19 acts as a HC gating modifier by interacting with the CT that forms a crucial gating element.https://www.mdpi.com/1422-0067/21/19/7340channel gatinggraphic user interfaceautomated analysistransition analysis |
| collection |
DOAJ |
| language |
English |
| format |
Article |
| sources |
DOAJ |
| author |
Alessio Lissoni Nan Wang Timur Nezlobinskii Maarten De Smet Alexander V. Panfilov Nele Vandersickel Luc Leybaert Katja Witschas |
| spellingShingle |
Alessio Lissoni Nan Wang Timur Nezlobinskii Maarten De Smet Alexander V. Panfilov Nele Vandersickel Luc Leybaert Katja Witschas Gap19, a Cx43 Hemichannel Inhibitor, Acts as a Gating Modifier That Decreases Main State Opening While Increasing Substate Gating International Journal of Molecular Sciences channel gating graphic user interface automated analysis transition analysis |
| author_facet |
Alessio Lissoni Nan Wang Timur Nezlobinskii Maarten De Smet Alexander V. Panfilov Nele Vandersickel Luc Leybaert Katja Witschas |
| author_sort |
Alessio Lissoni |
| title |
Gap19, a Cx43 Hemichannel Inhibitor, Acts as a Gating Modifier That Decreases Main State Opening While Increasing Substate Gating |
| title_short |
Gap19, a Cx43 Hemichannel Inhibitor, Acts as a Gating Modifier That Decreases Main State Opening While Increasing Substate Gating |
| title_full |
Gap19, a Cx43 Hemichannel Inhibitor, Acts as a Gating Modifier That Decreases Main State Opening While Increasing Substate Gating |
| title_fullStr |
Gap19, a Cx43 Hemichannel Inhibitor, Acts as a Gating Modifier That Decreases Main State Opening While Increasing Substate Gating |
| title_full_unstemmed |
Gap19, a Cx43 Hemichannel Inhibitor, Acts as a Gating Modifier That Decreases Main State Opening While Increasing Substate Gating |
| title_sort |
gap19, a cx43 hemichannel inhibitor, acts as a gating modifier that decreases main state opening while increasing substate gating |
| publisher |
MDPI AG |
| series |
International Journal of Molecular Sciences |
| issn |
1661-6596 1422-0067 |
| publishDate |
2020-10-01 |
| description |
Cx43 hemichannels (HCs) are electrically and chemically gated transmembrane pores with low open probability and multiple conductance states, which makes kinetic studies of channel gating in large datasets challenging. Here, we developed open access software, named HemiGUI, to analyze HC gating transitions and investigated voltage-induced HC opening based on up to ≈4000 events recorded in HeLa-Cx43-overexpressing cells. We performed a detailed characterization of Cx43 HC gating profiles and specifically focused on the role of the C-terminal tail (CT) domain by recording the impact of adding an EGFP tag to the Cx43 CT end (Cx43-EGFP) or by supplying the Cx43 HC-inhibiting peptide Gap19 that interferes with CT interaction with the cytoplasmic loop (CL). We found that Gap19 not only decreased HC opening activity to the open state (≈217 pS) but also increased the propensity of subconductance (≈80 pS) transitions that additionally became slower as compared to the control. The work demonstrates that large sample transition analysis allows detailed investigations on Cx43 HC gating and shows that Gap19 acts as a HC gating modifier by interacting with the CT that forms a crucial gating element. |
| topic |
channel gating graphic user interface automated analysis transition analysis |
| url |
https://www.mdpi.com/1422-0067/21/19/7340 |
| work_keys_str_mv |
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