Intersubunit concerted cooperative and cis-type mechanisms modulate allosteric gating in two-pore-domain potassium channel TREK-2
In response to diverse stimuli, two-pore-domain potassium channel TREK-2 regulates cellular excitability, and hence plays a key role in mediating neuropathic pain, mood disorders and ischemia through. Although more and more input modalities are found to achieve their modulations via acting on the ch...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2016-05-01
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| Series: | Frontiers in Cellular Neuroscience |
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| Online Access: | http://journal.frontiersin.org/Journal/10.3389/fncel.2016.00127/full |
| _version_ | 1818519129009684480 |
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| author | Ren-Gong eZhuo Peng ePeng Xiao-Yan eLiu Hai-Tao eYan Jiang-Ping eXu Jian-Quan eZheng Xiao-Li eWei Xiao-Yun eMa |
| author_facet | Ren-Gong eZhuo Peng ePeng Xiao-Yan eLiu Hai-Tao eYan Jiang-Ping eXu Jian-Quan eZheng Xiao-Li eWei Xiao-Yun eMa |
| author_sort | Ren-Gong eZhuo |
| collection | DOAJ |
| description | In response to diverse stimuli, two-pore-domain potassium channel TREK-2 regulates cellular excitability, and hence plays a key role in mediating neuropathic pain, mood disorders and ischemia through. Although more and more input modalities are found to achieve their modulations via acting on the channel, the potential role of subunit interaction in these modulations remains to be explored. In the current study, the deletion (lack of proximal C-terminus, pCt) or point mutation (G312A) was introduced into TREK-2 subunits to limit K+ conductance and used to report subunit stoichiometry. The constructs were then combined with wild type (WT) subunit to produce concatenated dimers with defined composition, and the gating kinetics of these channels to 2-Aminoethoxydiphenyl borate (2-APB) and extracellular pH (pHo) were characterized. Our results show that combination of WT and pCt/G312A subunits reserves similar gating properties to that of WT dimmers, suggesting that the WT subunit exerts dominant and positive effects on the mutated one, and thus the two subunits controls channel gating via a concerted cooperative manner. Further introduction of pCt into the latter subunit of heterodimeric channel G312A-WT or G312A-G312A attenuated their sensitivity to 2-APB and pHo alkalization, implicating that these signals were transduced by a cis-type mechanism. Together, our findings elucidate the mechanisms for how the two subunits control the pore gating of TREK-2, in which both intersubunit concerted cooperative and cis-type manners modulate the allosteric regulations induced by 2-APB and pHo alkalization. |
| first_indexed | 2024-12-11T01:19:36Z |
| format | Article |
| id | doaj.art-82379bd620c44f85805642abf9550c46 |
| institution | Directory Open Access Journal |
| issn | 1662-5102 |
| language | English |
| last_indexed | 2024-12-11T01:19:36Z |
| publishDate | 2016-05-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Cellular Neuroscience |
| spelling | doaj.art-82379bd620c44f85805642abf9550c462022-12-22T01:25:44ZengFrontiers Media S.A.Frontiers in Cellular Neuroscience1662-51022016-05-011010.3389/fncel.2016.00127188397Intersubunit concerted cooperative and cis-type mechanisms modulate allosteric gating in two-pore-domain potassium channel TREK-2Ren-Gong eZhuo0Peng ePeng1Xiao-Yan eLiu2Hai-Tao eYan3Jiang-Ping eXu4Jian-Quan eZheng5Xiao-Li eWei6Xiao-Yun eMa7Beijing Institute of Pharmacology and ToxicologyBeijing Institute of Pharmacology and ToxicologyBeijing Institute of Pharmacology and ToxicologyBeijing Institute of Pharmacology and ToxicologySouthern Medical UniversityBeijing Institute of Pharmacology and ToxicologyBeijing Institute of Pharmacology and ToxicologyBeijing Institute of Pharmacology and ToxicologyIn response to diverse stimuli, two-pore-domain potassium channel TREK-2 regulates cellular excitability, and hence plays a key role in mediating neuropathic pain, mood disorders and ischemia through. Although more and more input modalities are found to achieve their modulations via acting on the channel, the potential role of subunit interaction in these modulations remains to be explored. In the current study, the deletion (lack of proximal C-terminus, pCt) or point mutation (G312A) was introduced into TREK-2 subunits to limit K+ conductance and used to report subunit stoichiometry. The constructs were then combined with wild type (WT) subunit to produce concatenated dimers with defined composition, and the gating kinetics of these channels to 2-Aminoethoxydiphenyl borate (2-APB) and extracellular pH (pHo) were characterized. Our results show that combination of WT and pCt/G312A subunits reserves similar gating properties to that of WT dimmers, suggesting that the WT subunit exerts dominant and positive effects on the mutated one, and thus the two subunits controls channel gating via a concerted cooperative manner. Further introduction of pCt into the latter subunit of heterodimeric channel G312A-WT or G312A-G312A attenuated their sensitivity to 2-APB and pHo alkalization, implicating that these signals were transduced by a cis-type mechanism. Together, our findings elucidate the mechanisms for how the two subunits control the pore gating of TREK-2, in which both intersubunit concerted cooperative and cis-type manners modulate the allosteric regulations induced by 2-APB and pHo alkalization.http://journal.frontiersin.org/Journal/10.3389/fncel.2016.00127/fullAllosteric Regulation2-APBTwo-pore domain potassium channelTREK-2intersubunit interactionconcatenated dimer |
| spellingShingle | Ren-Gong eZhuo Peng ePeng Xiao-Yan eLiu Hai-Tao eYan Jiang-Ping eXu Jian-Quan eZheng Xiao-Li eWei Xiao-Yun eMa Intersubunit concerted cooperative and cis-type mechanisms modulate allosteric gating in two-pore-domain potassium channel TREK-2 Frontiers in Cellular Neuroscience Allosteric Regulation 2-APB Two-pore domain potassium channel TREK-2 intersubunit interaction concatenated dimer |
| title | Intersubunit concerted cooperative and cis-type mechanisms modulate allosteric gating in two-pore-domain potassium channel TREK-2 |
| title_full | Intersubunit concerted cooperative and cis-type mechanisms modulate allosteric gating in two-pore-domain potassium channel TREK-2 |
| title_fullStr | Intersubunit concerted cooperative and cis-type mechanisms modulate allosteric gating in two-pore-domain potassium channel TREK-2 |
| title_full_unstemmed | Intersubunit concerted cooperative and cis-type mechanisms modulate allosteric gating in two-pore-domain potassium channel TREK-2 |
| title_short | Intersubunit concerted cooperative and cis-type mechanisms modulate allosteric gating in two-pore-domain potassium channel TREK-2 |
| title_sort | intersubunit concerted cooperative and cis type mechanisms modulate allosteric gating in two pore domain potassium channel trek 2 |
| topic | Allosteric Regulation 2-APB Two-pore domain potassium channel TREK-2 intersubunit interaction concatenated dimer |
| url | http://journal.frontiersin.org/Journal/10.3389/fncel.2016.00127/full |
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