caa a22 4500 606175482 CHVBK 20210128100743.0 cr unu---uuuuu 210128e20150401xx s 000 0 eng 10.1007/s00018-014-1747-4 doi (NATIONALLICENCE)springer-10.1007/s00018-014-1747-4 Functional and pharmacological induced structural changes of the cystic fibrosis transmembrane conductance regulator in the membrane solved using SAXS [Elektronische Daten] [Debora Baroni, Olga Zegarra-Moran, Oscar Moran] The cystic fibrosis transmembrane conductance regulator (CFTR) chloride channel is a membrane-integral protein that belongs to the ATP-binding cassette superfamily. Mutations in the CFTR gene cause cystic fibrosis in which salt, water, and protein transports are defective in various tissues. To investigate the conformation of the CFTR in the membrane, we applied the small-angle x-ray scattering (SAXS) technique on microsomal membranes extracted from NIH/3T3 cells permanentely transfected with wild-type (WT) CFTR and with CFTR carrying the ΔF508 mutation. The electronic density profile of the membranes was calculated from the SAXS data, assuming the lipid bilayer electronic density to be composed by a series of Gaussian shells. The data indicate that membranes in the microsome vesicles, that contain mostly endoplasmic reticulum membranes, are oriented in the outside-out conformation. Phosphorylation does not change significantly the electronic density profile, while dephosphorylation produces a significant modification in the inner side of the profile. Thus, we conclude that the CFTR and its associated protein complex in microsomes are mostly phosphorylated. The electronic density profile of the ΔF508-CFTR microsomes is completely different from WT, suggesting a different assemblage of the proteins in the membranes. Low-temperature treatment of cells rescues the ΔF508-CFTR protein, resulting in a conformation that resembles the WT. Differently, treatment with the corrector VX-809 modifies the electronic profile of ΔF508-CFTR membrane, but does not recover completely the WT conformation. To our knowledge, this is the first report of a directphysical measurement of the structure of membranes containing CFTR in its native environment and in different functional and pharmacological conditions. Springer Basel, 2014 CFTR nationallicence Phosphorylation nationallicence Membrane structure nationallicence ΔF508 rescue nationallicence Cystic fibrosis nationallicence Small angle X-ray scattering nationallicence Baroni Debora Istituto di Biofisica, CNR, via De Marini, 6, 16149, Genoa, Italy aut Zegarra-Moran Olga Unità Operativa di Genetica Medica, Istituto G. Gaslini, Genoa, Italy aut Moran Oscar Istituto di Biofisica, CNR, via De Marini, 6, 16149, Genoa, Italy aut Cellular and Molecular Life Sciences Springer Basel 72/7(2015-04-01), 1363-1375 1420-682X 72:7<1363 2015 72 18 https://doi.org/10.1007/s00018-014-1747-4 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00018-014-1747-4 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Baroni Debora Istituto di Biofisica, CNR, via De Marini, 6, 16149, Genoa, Italy aut NATIONALLICENCE 700 1- Zegarra-Moran Olga Unità Operativa di Genetica Medica, Istituto G. Gaslini, Genoa, Italy aut NATIONALLICENCE 700 1- Moran Oscar Istituto di Biofisica, CNR, via De Marini, 6, 16149, Genoa, Italy aut NATIONALLICENCE 773 0- Cellular and Molecular Life Sciences Springer Basel 72/7(2015-04-01), 1363-1375 1420-682X 72:7<1363 2015 72 18