5JAE
LeuT in the outward-oriented, Na+-free return state, P21 form at pH 6.5
Summary for 5JAE
Entry DOI | 10.2210/pdb5jae/pdb |
Descriptor | Transporter, octyl beta-D-glucopyranoside (3 entities in total) |
Functional Keywords | membrane protein, neurotransmitter:sodium symporter family, amino acid transporter |
Biological source | Aquifex aeolicus (strain VF5) |
Total number of polymer chains | 2 |
Total formula weight | 119370.93 |
Authors | Malinauskaite, L.,Sahin, C.,Said, S.,Grouleff, J.,Shahsavar, A.,Bjerregaard, H.,Noer, P.,Severinsen, K.,Boesen, T.,Schiott, B.,Sinning, S.,Nissen, P. (deposition date: 2016-04-12, release date: 2016-06-01, Last modification date: 2024-01-10) |
Primary citation | Malinauskaite, L.,Said, S.,Sahin, C.,Grouleff, J.,Shahsavar, A.,Bjerregaard, H.,Noer, P.,Severinsen, K.,Boesen, T.,Schitt, B.,Sinning, S.,Nissen, P. A conserved leucine occupies the empty substrate site of LeuT in the Na(+)-free return state. Nat Commun, 7:11673-11673, 2016 Cited by PubMed Abstract: Bacterial members of the neurotransmitter:sodium symporter (NSS) family perform Na(+)-dependent amino-acid uptake and extrude H(+) in return. Previous NSS structures represent intermediates of Na(+)/substrate binding or intracellular release, but not the inward-to-outward return transition. Here we report crystal structures of Aquifex aeolicus LeuT in an outward-oriented, Na(+)- and substrate-free state likely to be H(+)-occluded. We find a remarkable rotation of the conserved Leu25 into the empty substrate-binding pocket and rearrangements of the empty Na(+) sites. Mutational studies of the equivalent Leu99 in the human serotonin transporter show a critical role of this residue on the transport rate. Molecular dynamics simulations show that extracellular Na(+) is blocked unless Leu25 is rotated out of the substrate-binding pocket. We propose that Leu25 facilitates the inward-to-outward transition by compensating a Na(+)- and substrate-free state and acts as the gatekeeper for Na(+) binding that prevents leak in inward-outward return transitions. PubMed: 27221344DOI: 10.1038/ncomms11673 PDB entries with the same primary citation |
Experimental method | X-RAY DIFFRACTION (2.5 Å) |
Structure validation
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