Refinement of the Central Steps of Substrate Transport by the Aspartate Transporter GltPh: Elucidating the Role of the Na2 Sodium Binding Site

• Published in: PLoS computational biology Vol. 11; no. 10; p. e1004551
• Main Authors: Venkatesan, SanthoshKannan, Saha, Kusumika, Sohail, Azmat, Sandtner, Walter, Freissmuth, Michael, Ecker, Gerhard F, Sitte, Harald H, Stockner, Thomas
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 01.10.2015; Public Library of Science (PLoS)
• Subjects: Alzheimers disease; Amino Acid Transport Systems - chemistry; Amino Acid Transport Systems - ultrastructure; Amyotrophic lateral sclerosis; Aspartate; Aspartic Acid - chemistry; Binding Sites; Chemical properties; Crystal structure; Diffusion; Experiments; Models, Chemical; Molecular Docking Simulation; Principal components analysis; Protein Binding; Protein Conformation; Simulation; Sodium; Sodium - chemistry; Structure
• ISSN: 1553-7358; 1553-734X; 1553-7358
• DOI: 10.1371/journal.pcbi.1004551

Glutamate homeostasis in the brain is maintained by glutamate transporter mediated accumulation. Impaired transport is associated with several neurological disorders, including stroke and amyotrophic lateral sclerosis. Crystal structures of the homolog transporter GltPh from Pyrococcus horikoshii revealed large structural changes. Substrate uptake at the atomic level and the mechanism of ion gradient conversion into directional transport remained enigmatic. We observed in repeated simulations that two local structural changes regulated transport. The first change led to formation of the transient Na2 sodium binding site, triggered by side chain rotation of T308. The second change destabilized cytoplasmic ionic interactions. We found that sodium binding to the transiently formed Na2 site energized substrate uptake through reshaping of the energy hypersurface. Uptake experiments in reconstituted proteoliposomes confirmed the proposed mechanism. We reproduced the results in the human glutamate transporter EAAT3 indicating a conserved mechanics from archaea to humans.