Transmembrane Helices 7 and 8 Confer Aggregation Sensitivity to the Cystic Fibrosis Transmembrane Conductance Regulator
The Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) is a large multi-spanning membrane protein that is susceptible to misfolding and aggregation. We have identified here the region responsible for this instability. Temperature-induced aggregation of C-terminally truncated versions of CFTR...
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MDPI AG
2023-10-01
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| Series: | International Journal of Molecular Sciences |
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| author | Bertrand Kleizen Eduardo de Mattos Olga Papaioannou Michele Monti Gian Gaetano Tartaglia Peter van der Sluijs Ineke Braakman |
| author_facet | Bertrand Kleizen Eduardo de Mattos Olga Papaioannou Michele Monti Gian Gaetano Tartaglia Peter van der Sluijs Ineke Braakman |
| author_sort | Bertrand Kleizen |
| collection | DOAJ |
| description | The Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) is a large multi-spanning membrane protein that is susceptible to misfolding and aggregation. We have identified here the region responsible for this instability. Temperature-induced aggregation of C-terminally truncated versions of CFTR demonstrated that all truncations up to the second transmembrane domain (TMD2), including the R region, largely resisted aggregation. Limited proteolysis identified a folded structure that was prone to aggregation and consisted of TMD2 and at least part of the Regulatory Region R. Only when both TM7 (TransMembrane helix 7) and TM8 were present, TMD2 fragments became as aggregation-sensitive as wild-type CFTR, in line with increased thermo-instability of late CFTR nascent chains and in silico prediction of aggregation propensity. In accord, isolated TMD2 was degraded faster in cells than isolated TMD1. We conclude that TMD2 extended at its N-terminus with part of the R region forms a protease-resistant structure that induces heat instability in CFTR and may be responsible for its limited intracellular stability. |
| first_indexed | 2024-03-11T11:28:32Z |
| format | Article |
| id | doaj.art-35296191939b461cb85b9dcd521552f2 |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-11T11:28:32Z |
| publishDate | 2023-10-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-35296191939b461cb85b9dcd521552f22023-11-10T15:05:08ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672023-10-0124211574110.3390/ijms242115741Transmembrane Helices 7 and 8 Confer Aggregation Sensitivity to the Cystic Fibrosis Transmembrane Conductance RegulatorBertrand Kleizen0Eduardo de Mattos1Olga Papaioannou2Michele Monti3Gian Gaetano Tartaglia4Peter van der Sluijs5Ineke Braakman6Cellular Protein Chemistry, Bijvoet Centre for Biomolecular Research, Utrecht University, 3584 CH Utrecht, The NetherlandsCellular Protein Chemistry, Bijvoet Centre for Biomolecular Research, Utrecht University, 3584 CH Utrecht, The NetherlandsCellular Protein Chemistry, Bijvoet Centre for Biomolecular Research, Utrecht University, 3584 CH Utrecht, The NetherlandsCenter for Life Nano- & Neuro-Science, Fondazione Istituto Italiano di Tecnologia (IIT), 00161 Rome, ItalyCenter for Life Nano- & Neuro-Science, Fondazione Istituto Italiano di Tecnologia (IIT), 00161 Rome, ItalyCellular Protein Chemistry, Bijvoet Centre for Biomolecular Research, Utrecht University, 3584 CH Utrecht, The NetherlandsCellular Protein Chemistry, Bijvoet Centre for Biomolecular Research, Utrecht University, 3584 CH Utrecht, The NetherlandsThe Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) is a large multi-spanning membrane protein that is susceptible to misfolding and aggregation. We have identified here the region responsible for this instability. Temperature-induced aggregation of C-terminally truncated versions of CFTR demonstrated that all truncations up to the second transmembrane domain (TMD2), including the R region, largely resisted aggregation. Limited proteolysis identified a folded structure that was prone to aggregation and consisted of TMD2 and at least part of the Regulatory Region R. Only when both TM7 (TransMembrane helix 7) and TM8 were present, TMD2 fragments became as aggregation-sensitive as wild-type CFTR, in line with increased thermo-instability of late CFTR nascent chains and in silico prediction of aggregation propensity. In accord, isolated TMD2 was degraded faster in cells than isolated TMD1. We conclude that TMD2 extended at its N-terminus with part of the R region forms a protease-resistant structure that induces heat instability in CFTR and may be responsible for its limited intracellular stability.https://www.mdpi.com/1422-0067/24/21/15741CFTRaggregationTMD2TM7TM8instability |
| spellingShingle | Bertrand Kleizen Eduardo de Mattos Olga Papaioannou Michele Monti Gian Gaetano Tartaglia Peter van der Sluijs Ineke Braakman Transmembrane Helices 7 and 8 Confer Aggregation Sensitivity to the Cystic Fibrosis Transmembrane Conductance Regulator International Journal of Molecular Sciences CFTR aggregation TMD2 TM7 TM8 instability |
| title | Transmembrane Helices 7 and 8 Confer Aggregation Sensitivity to the Cystic Fibrosis Transmembrane Conductance Regulator |
| title_full | Transmembrane Helices 7 and 8 Confer Aggregation Sensitivity to the Cystic Fibrosis Transmembrane Conductance Regulator |
| title_fullStr | Transmembrane Helices 7 and 8 Confer Aggregation Sensitivity to the Cystic Fibrosis Transmembrane Conductance Regulator |
| title_full_unstemmed | Transmembrane Helices 7 and 8 Confer Aggregation Sensitivity to the Cystic Fibrosis Transmembrane Conductance Regulator |
| title_short | Transmembrane Helices 7 and 8 Confer Aggregation Sensitivity to the Cystic Fibrosis Transmembrane Conductance Regulator |
| title_sort | transmembrane helices 7 and 8 confer aggregation sensitivity to the cystic fibrosis transmembrane conductance regulator |
| topic | CFTR aggregation TMD2 TM7 TM8 instability |
| url | https://www.mdpi.com/1422-0067/24/21/15741 |
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