@article{2af02bc688df44d2a9d12b0d5d8ace54,
title = "Structural basis for the reaction cycle of dass dicarboxylate transporters",
abstract = "Citrate, α-ketoglutarate and succinate are TCA cycle intermediates that also play essential roles in metabolic signaling and cellular regulation. These di-and tricarboxylates are imported into the cell by the divalent anion sodium symporter (DASS) family of plasma membrane transporters, which contains both cotransporters and exchangers. While DASS proteins transport substrates via an elevator mechanism, to date structures are only available for a single DASS cotransporter protein in a substrate-bound, inward-facing state. We report multiple cryo-EM and X-ray structures in four different states, including three hitherto unseen states, along with molecular dynamics simulations, of both a cotransporter and an exchanger. Comparison of these outward-and inward-facing structures reveal how the transport domain translates and rotates within the framework of the scaffold domain through the transport cycle. Additionally, we propose that DASS transporters ensure substrate coupling by a charge-compensation mechanism, and by structural changes upon substrate release.",
author = "Sauer, {David B.} and Noah Trebesch and Marden, {Jennifer J.} and Nicolette Cocco and Jinmei Song and Akiko Koide and Shohei Koide and Emad Tajkhorshid and Wang, {Da Neng}",
note = "Funding Information: is a DASS exchanger. This is supported by its phylogeny and ability to catalyze Funding Information: and XSEDE (TG-MCA06N060) to E.T. Blue Waters is supported by the NSF (OCI- Funding Information: (W81XWH-16-1-0153). N.T. was supported by a NSF Graduate Research Fellowship Funding Information: This work was financially supported by the NIH (R01NS108151, R01GM121994 and R01-DK099023 to D.N.W; U54GM095315 to W.A. Hendrickson; P41-GM104601 and R01-GM067887 to E.T.), the TESS Research Foundation and the American Epilepsy Society (to D.N.W). D.B.S. was supported by the American Cancer Society Postdoctoral Fellowship (129844-PF-17-135-01-TBE) and Department of Defense Horizon Award (W81XWH-16-1-0153). N.T. was supported by a NSF Graduate Research Fellowship (1746047). N.C. was supported by an NIH Predoctoral Training Grant (T32-GM088118). VcINDY homologs were cloned by B. Kloss at the New York Consortium of Membrane Protein Structures. We thank the following colleagues for reagent, technical assistance, and helpful discussions: J.P. Armache, J.G. Belasco, N. Coudray, T. Hattori, J. Jiang, N.K. Karpowich, M. Lopez Redondo, Z. Liu, R. Mancusso, A.B. Rejto and S.G. Sligar. We are also grateful to the staff at the following facilities for assistance in screening and data collection in cryo-EM and X-ray diffraction: the NYU Cryo-EM Facility, the Pacific Northwest Center for Cryo-EM, Advanced Light Source Beamline 5.0.2 at the Berkeley National Laboratory, beamlines AMX and FMX at NSLS-II and 19-BM and 19-ID at the Advanced Photon Source. EM data processing used computing resources at the HPC Facility of NYULMC, and we were assisted by A. Siavosh-Haghighi and M. Costantino. MD simulations were performed using supercomputing allocations from Blue Waters and XSEDE (TG-MCA06N060) to E.T. Blue Waters is supported by the NSF (OCI-0725070 and ACI-1238993), the State of Illinois and the National Geospatial-Intelligence Agency. XSEDE is supported by the NSF (ACI-1548562). Funding Information: (1746047). N.C. was supported by an NIH Predoctoral Training Grant (T32-GM088118). Funding Information: This work was financially supported by the NIH (R01NS108151, R01GM121994 and Publisher Copyright: {\textcopyright} 2020, eLife Sciences Publications Ltd. All rights reserved.",
year = "2020",
month = sep,
doi = "10.7554/ELIFE.61350",
language = "English (US)",
volume = "9",
pages = "1--74",
journal = "eLife",
issn = "2050-084X",
publisher = "eLife Sciences Publications",
}