Mutation-induced changes of transmembrane pore size revealed by combined ion-channel conductance and single vesicle permeabilization analyses
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Otros documentos de la autoría: Largo, Eneko; Gladue, Douglas; Torralba, Johana; Aguilella, Vicente; Alcaraz, Antonio; Borca, Manuel V.; Nieva, José L.
Metadatos
Mostrar el registro completo del ítemcomunitat-uji-handle:10234/9
comunitat-uji-handle2:10234/2507
comunitat-uji-handle3:10234/6973
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INVESTIGACIONMetadatos
Título
Mutation-induced changes of transmembrane pore size revealed by combined ion-channel conductance and single vesicle permeabilization analysesAutoría
Fecha de publicación
2018-05Editor
ElsevierISSN
0005-2736Cita bibliográfica
LARGO, Eneko, et al. Mutation-induced changes of transmembrane pore size revealed by combined ion-channel conductance and single vesicle permeabilization analyses. Biochimica et Biophysica Acta (BBA)-Biomembranes, 2018, vol. 1860, no 5, p. 1015-1021Tipo de documento
info:eu-repo/semantics/articleVersión de la editorial
https://www.sciencedirect.com/science/article/pii/S0005273618300129Versión
info:eu-repo/semantics/acceptedVersionPalabras clave / Materias
Resumen
Permeabilization of the Endoplasmic Reticulum (ER) is instrumental in the progression of host-cell infection by many viral pathogens. We have described that permeabilization of ER model membranes by the pore-forming ... [+]
Permeabilization of the Endoplasmic Reticulum (ER) is instrumental in the progression of host-cell infection by many viral pathogens. We have described that permeabilization of ER model membranes by the pore-forming domain of the Classical Swine Fever Virus (CSFV) p7 protein depends on two sequence determinants: the C-terminal transmembrane helix, and the preceding polar loop that regulates its activity. Here, by combining ion-channel activity measurements in planar lipid bilayers with imaging of single Giant Unilamellar Vesicles (GUVs), we demonstrate that point substitutions directed to conserved residues within these regions affect ER-like membrane permeabilization following distinct mechanisms. Whereas the polar loop appeared to be involved in protein insertion and oligomerization, substitution of residues predicted to face the lumen of the pore inhibited large conducting channels (>1 nS) over smaller ones (120 pS). Quantitative analyses of the ER-GUV distribution as a function of the solute size revealed a selective inhibition for the permeation of solutes with sizes larger than 4 kDa, further demonstrating that the mutation targeting the transmembrane helix prevented formation of the large pores. Collectively, our data support the idea that the pore-forming domain of p7 may assemble into finite pores with approximate diameters of 1 and 5 nm. Moreover, the observation that the mutation interfering with formation of the larger pores can hamper virus production without affecting ER localization or homo-oligomerization, suggests prospective strategies to block/attenuate pestiviruses. [-]
Publicado en
Biochimica et Biophysica Acta (BBA)-Biomembranes, 2018, vol. 1860, no 5Proyecto de investigación
USDA Agricultural Research Service (ARS-USDA): 8064-32000-056-18S; Basque Government: IT838-13; Ministry of Economy and Competitiveness of Spain: FIS2013-40473-P, FIS2016-75257-P; Universitat Jaume I: P1.1B2015-28Derechos de acceso
Copyright © Elsevier B.V.
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info:eu-repo/semantics/openAccess
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info:eu-repo/semantics/openAccess
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