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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.<\/jats:p>","DOI":"10.1073\/pnas.1115581109","type":"journal-article","created":{"date-parts":[[2012,1,10]],"date-time":"2012-01-10T01:43:26Z","timestamp":1326159806000},"update-policy":"https:\/\/doi.org\/10.1073\/pnas.cm10313","source":"Crossref","is-referenced-by-count":43,"title":["Mechanism of proton\/substrate coupling in the heptahelical lysosomal transporter cystinosin"],"prefix":"10.1073","volume":"109","author":[{"given":"Raquel","family":"Ruivo","sequence":"first","affiliation":[{"name":"Universit\u00e9 Paris Descartes, Sorbonne Paris Cit\u00e9, Centre National de la Recherche Scientifique, Unit\u00e9 Mixte de Recherche 8192, Centre Universitaire des Saints-P\u00e8res, 45 rue des Saints-P\u00e8res, F-75006 Paris, 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