Cardiac ryanodine receptor distribution is dynamic and changed by auxiliary proteins and post-translational modification
The effects of the immunophilins, FKBP12 and FKBP12.6, and phosphorylation on type II ryanodine receptor (RyR2) arrangement and function were examined using correlation microscopy (line scan confocal imaging of Ca2+ sparks and dual-tilt electron tomography) and dSTORM imaging of permeabilized Wistar...
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| Format: | Article |
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eLife Sciences Publications Ltd
2020-01-01
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| Series: | eLife |
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| Online Access: | https://elifesciences.org/articles/51602 |
| _version_ | 1811253276384952320 |
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| author | Parisa Asghari David RL Scriven Myles Ng Pankaj Panwar Keng C Chou Filip van Petegem Edwin DW Moore |
| author_facet | Parisa Asghari David RL Scriven Myles Ng Pankaj Panwar Keng C Chou Filip van Petegem Edwin DW Moore |
| author_sort | Parisa Asghari |
| collection | DOAJ |
| description | The effects of the immunophilins, FKBP12 and FKBP12.6, and phosphorylation on type II ryanodine receptor (RyR2) arrangement and function were examined using correlation microscopy (line scan confocal imaging of Ca2+ sparks and dual-tilt electron tomography) and dSTORM imaging of permeabilized Wistar rat ventricular myocytes. Saturating concentrations (10 µmol/L) of either FKBP12 or 12.6 significantly reduced the frequency, spread, amplitude and Ca2+ spark mass relative to control, while the tomograms revealed both proteins shifted the tetramers into a largely side-by-side configuration. Phosphorylation of immunophilin-saturated RyR2 resulted in structural and functional changes largely comparable to phosphorylation alone. dSTORM images of myocyte surfaces demonstrated that both FKBP12 and 12.6 significantly reduced RyR2 cluster sizes, while phosphorylation, even of immunophilin-saturated RyR2, increased them. We conclude that both RyR2 cluster size and the arrangement of tetramers within clusters is dynamic and respond to changes in the cellular environment. Further, these changes affect Ca2+ spark formation. |
| first_indexed | 2024-04-12T16:47:29Z |
| format | Article |
| id | doaj.art-da3ac013f2e848feb2dbf8e6e0317711 |
| institution | Directory Open Access Journal |
| issn | 2050-084X |
| language | English |
| last_indexed | 2024-04-12T16:47:29Z |
| publishDate | 2020-01-01 |
| publisher | eLife Sciences Publications Ltd |
| record_format | Article |
| series | eLife |
| spelling | doaj.art-da3ac013f2e848feb2dbf8e6e03177112022-12-22T03:24:31ZengeLife Sciences Publications LtdeLife2050-084X2020-01-01910.7554/eLife.51602Cardiac ryanodine receptor distribution is dynamic and changed by auxiliary proteins and post-translational modificationParisa Asghari0https://orcid.org/0000-0003-2285-3242David RL Scriven1https://orcid.org/0000-0003-1828-1405Myles Ng2Pankaj Panwar3Keng C Chou4Filip van Petegem5https://orcid.org/0000-0003-2728-8537Edwin DW Moore6https://orcid.org/0000-0001-7519-5592Department of Cellular and Physiological Sciences, University of British Columbia, Vancouver, CanadaDepartment of Cellular and Physiological Sciences, University of British Columbia, Vancouver, CanadaDepartment of Cellular and Physiological Sciences, University of British Columbia, Vancouver, CanadaDepartment of Biochemistry and Molecular Biology, University of British Columbia, Vancouver, CanadaDepartment of Chemistry, University of British Columbia, Vancouver, CanadaDepartment of Biochemistry and Molecular Biology, University of British Columbia, Vancouver, CanadaDepartment of Cellular and Physiological Sciences, University of British Columbia, Vancouver, CanadaThe effects of the immunophilins, FKBP12 and FKBP12.6, and phosphorylation on type II ryanodine receptor (RyR2) arrangement and function were examined using correlation microscopy (line scan confocal imaging of Ca2+ sparks and dual-tilt electron tomography) and dSTORM imaging of permeabilized Wistar rat ventricular myocytes. Saturating concentrations (10 µmol/L) of either FKBP12 or 12.6 significantly reduced the frequency, spread, amplitude and Ca2+ spark mass relative to control, while the tomograms revealed both proteins shifted the tetramers into a largely side-by-side configuration. Phosphorylation of immunophilin-saturated RyR2 resulted in structural and functional changes largely comparable to phosphorylation alone. dSTORM images of myocyte surfaces demonstrated that both FKBP12 and 12.6 significantly reduced RyR2 cluster sizes, while phosphorylation, even of immunophilin-saturated RyR2, increased them. We conclude that both RyR2 cluster size and the arrangement of tetramers within clusters is dynamic and respond to changes in the cellular environment. Further, these changes affect Ca2+ spark formation.https://elifesciences.org/articles/51602ryanodine receptorexcitation-contraction couplingimmunophilins3D electron tomographysuperresolution microscopy |
| spellingShingle | Parisa Asghari David RL Scriven Myles Ng Pankaj Panwar Keng C Chou Filip van Petegem Edwin DW Moore Cardiac ryanodine receptor distribution is dynamic and changed by auxiliary proteins and post-translational modification eLife ryanodine receptor excitation-contraction coupling immunophilins 3D electron tomography superresolution microscopy |
| title | Cardiac ryanodine receptor distribution is dynamic and changed by auxiliary proteins and post-translational modification |
| title_full | Cardiac ryanodine receptor distribution is dynamic and changed by auxiliary proteins and post-translational modification |
| title_fullStr | Cardiac ryanodine receptor distribution is dynamic and changed by auxiliary proteins and post-translational modification |
| title_full_unstemmed | Cardiac ryanodine receptor distribution is dynamic and changed by auxiliary proteins and post-translational modification |
| title_short | Cardiac ryanodine receptor distribution is dynamic and changed by auxiliary proteins and post-translational modification |
| title_sort | cardiac ryanodine receptor distribution is dynamic and changed by auxiliary proteins and post translational modification |
| topic | ryanodine receptor excitation-contraction coupling immunophilins 3D electron tomography superresolution microscopy |
| url | https://elifesciences.org/articles/51602 |
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