| Autor: |
Chi Wang, Zhengrong Yang, Blaine J. Loughlin, Haijin Xu, Guido Veit, Sergey Vorobiev, Oliver B. Clarke, Fan Jiang, Yaohui Li, Shikha Singh, Zachary Rich, Elizabeth R. Menten, Robert A. Grassucci, Wei Wang, Allison Mezzell, Ziao Fu, Kam-Ho Wong, Jing Wang, Diana R. Wetmore, R. Bryan Sutton, Christie G. Brouillette, Ina L. Urbatsch, John C. Kappes, Gergely L. Lukacs, Joachim Frank, John F. Hunt |
| Rok vydání: |
2022 |
| DOI: |
10.1101/2022.10.10.510913 |
| Popis: |
Cystic fibrosis (CF) is caused by mutations in a chloride channel called the human Cystic Fibrosis Transmembrane Conductance Regulator (hCFTR). We used cryo-EM global conformational ensemble reconstruction to characterize the mechanism by which the breakthrough drug VX445 (Elexacaftor) simultaneously corrects both protein-folding and channel-gating defects caused by CF mutations. VX445 drives hCFTR molecules harboring the gating-defective G551D mutation towards the open-channel conformation by binding to a site in the first transmembrane domain. This binding interaction reverses the usual pathway of allosteric structural communication by which ATP binding activates channel conductance, which is blocked by the G551D mutation. Our ensemble reconstructions include a 3.4 Å non-native structure demonstrating that detachment of the first nucleotide-binding domain of hCFTR is directly coupled to local unfolding of the VX445 binding site. Reversal of this unfolding transition likely contributes to its corrector activity by cooperatively stabilizing NBD1 and the transmembrane domains of hCFTR during biogenesis.SummaryCryo-EM global conformational ensemble reconstruction has been used to characterize the mechanism-of-action of a breakthrough pharmaceutical that corrects fatal protein-folding and channel-gating defects in the human cystic fibrosis transmembrane conductance regulator (CFTR). |
| Databáze: |
OpenAIRE |
| Externí odkaz: |
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