The Human Acid-Sensing Ion Channel ASIC1a: Evidence for a Homotetrameric Assembly State at the Cell Surface.
The chicken acid-sensing ion channel ASIC1 has been crystallized as a homotrimer. We address here the oligomeric state of the functional ASIC1 in situ at the cell surface. The oligomeric states of functional ASIC1a and mutants with additional cysteines introduced in the extracellular pore vestibule...
| Main Authors: | , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2015-01-01
|
| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC4529235?pdf=render |
| _version_ | 1818848112715759616 |
|---|---|
| author | Miguel Xavier van Bemmelen Delphine Huser Ivan Gautschi Laurent Schild |
| author_facet | Miguel Xavier van Bemmelen Delphine Huser Ivan Gautschi Laurent Schild |
| author_sort | Miguel Xavier van Bemmelen |
| collection | DOAJ |
| description | The chicken acid-sensing ion channel ASIC1 has been crystallized as a homotrimer. We address here the oligomeric state of the functional ASIC1 in situ at the cell surface. The oligomeric states of functional ASIC1a and mutants with additional cysteines introduced in the extracellular pore vestibule were resolved on SDS-PAGE. The functional ASIC1 complexes were stabilized at the cell surface of Xenopus laevis oocytes or CHO cells either using the sulfhydryl crosslinker BMOE, or sodium tetrathionate (NaTT). Under these different crosslinking conditions ASIC1a migrates as four distinct oligomeric states that correspond by mass to multiples of a single ASIC1a subunit. The relative importance of each of the four ASIC1a oligomers was critically dependent on the availability of cysteines in the transmembrane domain for crosslinking, consistent with the presence of ASIC1a homo-oligomers. The expression of ASIC1a monomers, trimeric or tetrameric concatemeric cDNA constructs resulted in functional channels. The resulting ASIC1a complexes are resolved as a predominant tetramer over the other oligomeric forms, after stabilization with BMOE or NaTT and SDS-PAGE/western blot analysis. Our data identify a major ASIC1a homotetramer at the surface membrane of the cell expressing functional ASIC1a channel. |
| first_indexed | 2024-12-19T06:12:10Z |
| format | Article |
| id | doaj.art-15b7c4e0f6b7422187e470be9dc57879 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-19T06:12:10Z |
| publishDate | 2015-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-15b7c4e0f6b7422187e470be9dc578792022-12-21T20:32:58ZengPublic Library of Science (PLoS)PLoS ONE1932-62032015-01-01108e013519110.1371/journal.pone.0135191The Human Acid-Sensing Ion Channel ASIC1a: Evidence for a Homotetrameric Assembly State at the Cell Surface.Miguel Xavier van BemmelenDelphine HuserIvan GautschiLaurent SchildThe chicken acid-sensing ion channel ASIC1 has been crystallized as a homotrimer. We address here the oligomeric state of the functional ASIC1 in situ at the cell surface. The oligomeric states of functional ASIC1a and mutants with additional cysteines introduced in the extracellular pore vestibule were resolved on SDS-PAGE. The functional ASIC1 complexes were stabilized at the cell surface of Xenopus laevis oocytes or CHO cells either using the sulfhydryl crosslinker BMOE, or sodium tetrathionate (NaTT). Under these different crosslinking conditions ASIC1a migrates as four distinct oligomeric states that correspond by mass to multiples of a single ASIC1a subunit. The relative importance of each of the four ASIC1a oligomers was critically dependent on the availability of cysteines in the transmembrane domain for crosslinking, consistent with the presence of ASIC1a homo-oligomers. The expression of ASIC1a monomers, trimeric or tetrameric concatemeric cDNA constructs resulted in functional channels. The resulting ASIC1a complexes are resolved as a predominant tetramer over the other oligomeric forms, after stabilization with BMOE or NaTT and SDS-PAGE/western blot analysis. Our data identify a major ASIC1a homotetramer at the surface membrane of the cell expressing functional ASIC1a channel.http://europepmc.org/articles/PMC4529235?pdf=render |
| spellingShingle | Miguel Xavier van Bemmelen Delphine Huser Ivan Gautschi Laurent Schild The Human Acid-Sensing Ion Channel ASIC1a: Evidence for a Homotetrameric Assembly State at the Cell Surface. PLoS ONE |
| title | The Human Acid-Sensing Ion Channel ASIC1a: Evidence for a Homotetrameric Assembly State at the Cell Surface. |
| title_full | The Human Acid-Sensing Ion Channel ASIC1a: Evidence for a Homotetrameric Assembly State at the Cell Surface. |
| title_fullStr | The Human Acid-Sensing Ion Channel ASIC1a: Evidence for a Homotetrameric Assembly State at the Cell Surface. |
| title_full_unstemmed | The Human Acid-Sensing Ion Channel ASIC1a: Evidence for a Homotetrameric Assembly State at the Cell Surface. |
| title_short | The Human Acid-Sensing Ion Channel ASIC1a: Evidence for a Homotetrameric Assembly State at the Cell Surface. |
| title_sort | human acid sensing ion channel asic1a evidence for a homotetrameric assembly state at the cell surface |
| url | http://europepmc.org/articles/PMC4529235?pdf=render |
| work_keys_str_mv | AT miguelxaviervanbemmelen thehumanacidsensingionchannelasic1aevidenceforahomotetramericassemblystateatthecellsurface AT delphinehuser thehumanacidsensingionchannelasic1aevidenceforahomotetramericassemblystateatthecellsurface AT ivangautschi thehumanacidsensingionchannelasic1aevidenceforahomotetramericassemblystateatthecellsurface AT laurentschild thehumanacidsensingionchannelasic1aevidenceforahomotetramericassemblystateatthecellsurface AT miguelxaviervanbemmelen humanacidsensingionchannelasic1aevidenceforahomotetramericassemblystateatthecellsurface AT delphinehuser humanacidsensingionchannelasic1aevidenceforahomotetramericassemblystateatthecellsurface AT ivangautschi humanacidsensingionchannelasic1aevidenceforahomotetramericassemblystateatthecellsurface AT laurentschild humanacidsensingionchannelasic1aevidenceforahomotetramericassemblystateatthecellsurface |