AutoriRuivo R, Bellenchi GC, Chen X, Zifarelli G, Sagne' C, Debacker C, Pusch M, Supplisson S, Gasnier B.
AbstractSecondary active transporters use electrochemical gradients provided by primary ion pumps to translocate metabolites or drugs "uphill" across membranes. Here we report the ion-coupling mechanism of cystinosin, an unusual eukaryotic, proton-driven transporter distantly related to the proton pump bacteriorhodopsin. In humans, cystinosin exports the proteolysis-derived dimeric amino acid cystine from lysosomes and is impaired in cystinosis. Using voltage-dependence analysis of steady-state and transient currents elicited by cystine and neutralization-scanning mutagenesis of conserved protonatable residues, we show that cystine binding is coupled to protonation of a clinically relevant aspartate buried in the membrane. Deuterium isotope substitution experiments are consistent with an access of this aspartate from the lysosomal lumen through a deep proton channel. This aspartate lies in one of the two PQ-loop motifs shared by cystinosin with a set of eukaryotic membrane proteins of unknown function and is conserved in about half of them, thus suggesting that other PQ-loop proteins may translocate protons.
RivistaProceedings Of The National Academy Of Sciences Of The United States Of America
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Pagina inizio217
Pagina fineE210
Autori IBFMichael PUSCH, Giovanni ZIFARELLI
Linee di Ricerca IBFMD.P01.009.001