Publication
Inhibition of calpain 1 restores plasma membrane stability to pharmacologically rescued Phe508del-CFTR variant
| dc.contributor.author | Matos, Ana M. | |
| dc.contributor.author | Pinto, Francisco R. | |
| dc.contributor.author | Barros, Patrícia | |
| dc.contributor.author | Amaral, Margarida D. | |
| dc.contributor.author | Pepperkok, Rainer | |
| dc.contributor.author | Matos, Paulo | |
| dc.date.accessioned | 2020-04-27T07:48:09Z | |
| dc.date.available | 2020-07-19T00:30:14Z | |
| dc.date.issued | 2019-09-06 | |
| dc.description.abstract | Cystic fibrosis (CF) is a genetic disease caused by mutations in the gene encoding CF transmembrane conductance regulator (CFTR), a chloride channel normally expressed at the surface of epithelial cells. The most frequent mutation, resulting in Phe-508 deletion, causes CFTR misfolding and its premature degradation. Low temperature or pharmacological correctors can partly rescue the Phe508del-CFTR processing defect and enhance trafficking of this channel variant to the plasma membrane (PM). Nevertheless, the rescued channels have an increased endocytosis rate, being quickly removed from the PM by the peripheral protein quality-control pathway. We previously reported that rescued Phe508del-CFTR (rPhe508del) can be retained at the cell surface by stimulating signaling pathways that coax the adaptor molecule ezrin (EZR) to tether rPhe508del-Na+/H+-exchange regulatory factor-1 complexes to the actin cytoskeleton, thereby averting the rapid internalization of this channel variant. However, the molecular basis for why rPhe508del fails to recruit active EZR to the PM remains elusive. Here, using a proteomics approach, we characterized and compared the core components of wt-CFTR- or rPhe508del-containing macromolecular complexes at the surface of human bronchial epithelial cells. We identified calpain 1 (CAPN1) as an exclusive rPhe508del interactor that prevents active EZR recruitment, impairs rPhe508del anchoring to actin, and reduces its stability in the PM. We show that either CAPN1 down-regulation or its chemical inhibition dramatically improves the functional rescue of Phe508del-CFTR in airway cells. These observations suggest that CAPN1 constitutes an appealing target for pharmacological intervention, as part of CF combination therapies restoring Phe508del-CFTR function. | pt_PT |
| dc.description.sponsorship | Work supported by grants PTDC/BIACEL/28408/2017 and IF2012 to PM and center grant UID/MULTI/04046/2019 to BioISI, both from FCT, Portugal. AMM is recipient of fellowship SFRH/BD/52490/2014 from BioSYS PhD programme PD65-2012, and PB of fellowship SFRH/BPD/94322/2013, both funded by FCT, Portugal. We would like to acknowledge Dr. P. Haggie, University of California, San Francisco [UCSF], School of Medicine for kind gift of halide-sensitive YFPF46L/H148Q/I152L (HS-YFP) plasmid. The authors declare no competing financial interests. | pt_PT |
| dc.description.version | info:eu-repo/semantics/publishedVersion | pt_PT |
| dc.identifier.citation | J Biol Chem. 2019 Sep 6;294(36):13396-13410. doi: 10.1074/jbc.RA119.008738. Epub 2019 Jul 19 | pt_PT |
| dc.identifier.doi | 10.1074/jbc.RA119.008738 | pt_PT |
| dc.identifier.issn | 0021-9258 | |
| dc.identifier.uri | http://hdl.handle.net/10400.18/6529 | |
| dc.language.iso | eng | pt_PT |
| dc.peerreviewed | yes | pt_PT |
| dc.publisher | American Society for Biochemistry and Molecular Biology | pt_PT |
| dc.relation.publisherversion | https://www.jbc.org/content/294/36/13396.long | pt_PT |
| dc.subject | Cystic Fibrosis | pt_PT |
| dc.subject | CFTR | pt_PT |
| dc.subject | Calpain-1 | pt_PT |
| dc.subject | Vias de Transdução de Sinal e Patologias Associadas | pt_PT |
| dc.title | Inhibition of calpain 1 restores plasma membrane stability to pharmacologically rescued Phe508del-CFTR variant | pt_PT |
| dc.type | journal article | |
| dspace.entity.type | Publication | |
| oaire.citation.endPage | 13410 | pt_PT |
| oaire.citation.issue | 36 | pt_PT |
| oaire.citation.startPage | 13396 | pt_PT |
| oaire.citation.title | Journal of Biological Chemistry | pt_PT |
| oaire.citation.volume | 294 | pt_PT |
| rcaap.embargofct | De acordo com política editorial da revista. | pt_PT |
| rcaap.rights | openAccess | pt_PT |
| rcaap.type | article | pt_PT |