Proximity Profiling of the CFTR Interaction Landscape in Response to Orkambi
Deletion of phenylalanine 508 (∆F508) of the Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) anion channel protein is the leading cause of Cystic Fibrosis (CF). Here, we report the analysis of CFTR and ∆F508-CFTR interactomes using BioID (proximity-dependent biotin identification), a tech...
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| Format: | Article |
| Language: | English |
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MDPI AG
2022-02-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/23/5/2442 |
| _version_ | 1797475009927577600 |
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| author | Melissa Iazzi Audrey Astori Jonathan St-Germain Brian Raught Gagan D. Gupta |
| author_facet | Melissa Iazzi Audrey Astori Jonathan St-Germain Brian Raught Gagan D. Gupta |
| author_sort | Melissa Iazzi |
| collection | DOAJ |
| description | Deletion of phenylalanine 508 (∆F508) of the Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) anion channel protein is the leading cause of Cystic Fibrosis (CF). Here, we report the analysis of CFTR and ∆F508-CFTR interactomes using BioID (proximity-dependent biotin identification), a technique that can also detect transient associations. We identified 474 high-confidence CFTR proximity-interactors, 57 of which have been previously validated, with the remainder representing novel interaction space. The ∆F508 interactome, comprising 626 proximity-interactors was markedly different from its wild type counterpart, with numerous alterations in protein associations categorized in membrane trafficking and cellular stress functions. Furthermore, analysis of the ∆F508 interactome in cells treated with Orkambi identified several interactions that were altered as a result of this drug therapy. We examined two candidate CFTR proximity interactors, VAPB and NOS1AP, in functional assays designed to assess surface delivery and overall chloride efflux. VAPB depletion impacted both CFTR surface delivery and chloride efflux, whereas NOS1AP depletion only affected the latter. The wild type and ∆F508-CFTR interactomes represent rich datasets that could be further mined to reveal additional candidates for the functional rescue of ∆F508-CFTR. |
| first_indexed | 2024-03-09T20:39:06Z |
| format | Article |
| id | doaj.art-943111a8614c4316865de90b53f34468 |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-09T20:39:06Z |
| publishDate | 2022-02-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-943111a8614c4316865de90b53f344682023-11-23T23:04:03ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672022-02-01235244210.3390/ijms23052442Proximity Profiling of the CFTR Interaction Landscape in Response to OrkambiMelissa Iazzi0Audrey Astori1Jonathan St-Germain2Brian Raught3Gagan D. Gupta4Department of Chemistry and Biology, Ryerson University, Toronto, ON M5B 2K3, CanadaDepartment of Medical Biophysics, University of Toronto, Toronto, ON M5S 1A1, CanadaDepartment of Medical Biophysics, University of Toronto, Toronto, ON M5S 1A1, CanadaDepartment of Medical Biophysics, University of Toronto, Toronto, ON M5S 1A1, CanadaDepartment of Chemistry and Biology, Ryerson University, Toronto, ON M5B 2K3, CanadaDeletion of phenylalanine 508 (∆F508) of the Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) anion channel protein is the leading cause of Cystic Fibrosis (CF). Here, we report the analysis of CFTR and ∆F508-CFTR interactomes using BioID (proximity-dependent biotin identification), a technique that can also detect transient associations. We identified 474 high-confidence CFTR proximity-interactors, 57 of which have been previously validated, with the remainder representing novel interaction space. The ∆F508 interactome, comprising 626 proximity-interactors was markedly different from its wild type counterpart, with numerous alterations in protein associations categorized in membrane trafficking and cellular stress functions. Furthermore, analysis of the ∆F508 interactome in cells treated with Orkambi identified several interactions that were altered as a result of this drug therapy. We examined two candidate CFTR proximity interactors, VAPB and NOS1AP, in functional assays designed to assess surface delivery and overall chloride efflux. VAPB depletion impacted both CFTR surface delivery and chloride efflux, whereas NOS1AP depletion only affected the latter. The wild type and ∆F508-CFTR interactomes represent rich datasets that could be further mined to reveal additional candidates for the functional rescue of ∆F508-CFTR.https://www.mdpi.com/1422-0067/23/5/2442CFTR interactionsCFTR modulatorscystic fibrosisOrkambitheratypingprotein trafficking |
| spellingShingle | Melissa Iazzi Audrey Astori Jonathan St-Germain Brian Raught Gagan D. Gupta Proximity Profiling of the CFTR Interaction Landscape in Response to Orkambi International Journal of Molecular Sciences CFTR interactions CFTR modulators cystic fibrosis Orkambi theratyping protein trafficking |
| title | Proximity Profiling of the CFTR Interaction Landscape in Response to Orkambi |
| title_full | Proximity Profiling of the CFTR Interaction Landscape in Response to Orkambi |
| title_fullStr | Proximity Profiling of the CFTR Interaction Landscape in Response to Orkambi |
| title_full_unstemmed | Proximity Profiling of the CFTR Interaction Landscape in Response to Orkambi |
| title_short | Proximity Profiling of the CFTR Interaction Landscape in Response to Orkambi |
| title_sort | proximity profiling of the cftr interaction landscape in response to orkambi |
| topic | CFTR interactions CFTR modulators cystic fibrosis Orkambi theratyping protein trafficking |
| url | https://www.mdpi.com/1422-0067/23/5/2442 |
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