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Conformational transitions of the sodium-dependent sugar transporter, vSGLT

Sodium-dependent transporters couple the flow of Na(+) ions down their electrochemical potential gradient to the uphill transport of various ligands. Many of these transporters share a common core structure composed of a five-helix inverted repeat and deliver their cargo utilizing an alternating-acc...

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Detalles Bibliográficos
Autores principales: Paz, Aviv, Claxton, Derek P., Kumar, Jay Prakash, Kazmier, Kelli, Bisignano, Paola, Sharma, Shruti, Nolte, Shannon A., Liwag, Terrin M., Nayak, Vinod, Wright, Ernest M., Grabe, Michael, Mchaourab, Hassane S., Abramson, Jeff
Formato: Online Artículo Texto
Lenguaje:English
Publicado: National Academy of Sciences 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5866573/
https://www.ncbi.nlm.nih.gov/pubmed/29507231
http://dx.doi.org/10.1073/pnas.1718451115
Descripción
Sumario:Sodium-dependent transporters couple the flow of Na(+) ions down their electrochemical potential gradient to the uphill transport of various ligands. Many of these transporters share a common core structure composed of a five-helix inverted repeat and deliver their cargo utilizing an alternating-access mechanism. A detailed characterization of inward-facing conformations of the Na(+)-dependent sugar transporter from Vibrio parahaemolyticus (vSGLT) has previously been reported, but structural details on additional conformations and on how Na(+) and ligand influence the equilibrium between other states remains unknown. Here, double electron–electron resonance spectroscopy, structural modeling, and molecular dynamics are utilized to deduce ligand-dependent equilibria shifts of vSGLT in micelles. In the absence and presence of saturating amounts of Na(+), vSGLT favors an inward-facing conformation. Upon binding both Na(+) and sugar, the equilibrium shifts toward either an outward-facing or occluded conformation. While Na(+) alone does not stabilize the outward-facing state, gating charge calculations together with a kinetic model of transport suggest that the resting negative membrane potential of the cell, absent in detergent-solubilized samples, may stabilize vSGLT in an outward-open conformation where it is poised for binding external sugars. In total, these findings provide insights into ligand-induced conformational selection and delineate the transport cycle of vSGLT.