Cryo-EM structure of the human Asc-1 transporter complex.
Journal
Nature communications
ISSN: 2041-1723
Titre abrégé: Nat Commun
Pays: England
ID NLM: 101528555
Informations de publication
Date de publication:
08 Apr 2024
08 Apr 2024
Historique:
received:
10
10
2023
accepted:
02
04
2024
medline:
9
4
2024
pubmed:
9
4
2024
entrez:
8
4
2024
Statut:
epublish
Résumé
The Alanine-Serine-Cysteine transporter 1 (Asc-1 or SLC7A10) forms a crucial heterodimeric transporter complex with 4F2hc (SLC3A2) through a covalent disulfide bridge. This complex enables the sodium-independent transport of small neutral amino acids, including L-Alanine (L-Ala), Glycine (Gly), and D-Serine (D-Ser), within the central nervous system (CNS). D-Ser and Gly are two key endogenous glutamate co-agonists that activate N-methyl-d-aspartate (NMDA) receptors by binding to the allosteric site. Mice deficient in Asc-1 display severe symptoms such as tremors, ataxia, and seizures, leading to early postnatal death. Despite its physiological importance, the functional mechanism of the Asc-1-4F2hc complex has remained elusive. Here, we present cryo-electron microscopy (cryo-EM) structures of the human Asc-1-4F2hc complex in its apo state, D-Ser bound state, and L-Ala bound state, resolved at 3.6 Å, 3.5 Å, and 3.4 Å, respectively. Through detailed structural analysis and transport assays, we uncover a comprehensive alternating access mechanism that underlies conformational changes in the complex. In summary, our findings reveal the architecture of the Asc-1 and 4F2hc complex and provide valuable insights into substrate recognition and the functional cycle of this essential transporter complex.
Identifiants
pubmed: 38589439
doi: 10.1038/s41467-024-47468-1
pii: 10.1038/s41467-024-47468-1
doi:
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
3036Informations de copyright
© 2024. The Author(s).
Références
Broer, S. & Palacin, M. The role of amino acid transporters in inherited and acquired diseases. Biochem. J. 436, 193–211 (2011).
pubmed: 21568940
doi: 10.1042/BJ20101912
Yahyaoui, R. & Perez-Frias, J. Amino acid transport defects in human inherited metabolic disorders. Int. J. Mol. Sci. 21, 119 (2019).
pubmed: 31878022
pmcid: 6981491
doi: 10.3390/ijms21010119
Verrey, F. et al. CATs and HATs: the SLC7 family of amino acid transporters. Pflug. Arch. 447, 532–542 (2004).
doi: 10.1007/s00424-003-1086-z
Pfeiffer, R. et al. Functional heterodimeric amino acid transporters lacking cysteine residues involved in disulfide bond. FEBS Lett. 439, 157–162 (1998).
pubmed: 9849898
doi: 10.1016/S0014-5793(98)01359-3
Fotiadis, D., Kanai, Y. & Palacin, M. The SLC3 and SLC7 families of amino acid transporters. Mol. Aspects Med. 34, 139–158 (2013).
pubmed: 23506863
doi: 10.1016/j.mam.2012.10.007
Mastroberardino, L. et al. Amino-acid transport by heterodimers of 4F2hc/CD98 and members of a permease family. Nature 395, 288–291 (1998).
pubmed: 9751058
doi: 10.1038/26246
Yan, R., Zhao, X., Lei, J. & Zhou, Q. Structure of the human LAT1-4F2hc heteromeric amino acid transporter complex. Nature 568, 127–130 (2019).
pubmed: 30867591
doi: 10.1038/s41586-019-1011-z
Kanai, Y. et al. Expression cloning and characterization of a transporter for large neutral amino acids activated by the heavy chain of 4F2 antigen (CD98). J. Biol. Chem. 273, 23629–23632 (1998).
pubmed: 9726963
doi: 10.1074/jbc.273.37.23629
Kim, D. K. et al. Expression of L-type amino acid transporter 1 (LAT1) and 4F2 heavy chain (4F2hc) in oral squamous cell carcinoma and its precusor lesions. Anticancer Res. 24, 1671–1675 (2004).
pubmed: 15274339
Toyoda, M. et al. Prognostic significance of amino-acid transporter expression (LAT1, ASCT2, and xCT) in surgically resected tongue cancer. Br. J. Cancer 110, 2506–2513 (2014).
pubmed: 24762957
pmcid: 4021522
doi: 10.1038/bjc.2014.178
Kaira, K. et al. l-type amino acid transporter 1 and CD98 expression in primary and metastatic sites of human neoplasms. Cancer Sci. 99, 2380–2386 (2008).
pubmed: 19018776
doi: 10.1111/j.1349-7006.2008.00969.x
Feliubadalo, L. et al. Non-type I cystinuria caused by mutations in SLC7A9, encoding a subunit (bo,+AT) of rBAT. Nat Genet 23, 52–57 (1999).
pubmed: 10471498
doi: 10.1038/12652
Torrents, D. et al. Identification and characterization of a membrane protein (y+L amino acid transporter-1) that associates with 4F2hc to encode the amino acid transport activity y+L. A candidate gene for lysinuric protein intolerance. J. Biol. Chem. 273, 32437–32445 (1998).
pubmed: 9829974
doi: 10.1074/jbc.273.49.32437
Yanagida, O. et al. Human L-type amino acid transporter 1 (LAT1): characterization of function and expression in tumor cell lines. Biochim. Biophys. Acta 1514, 291–302 (2001).
pubmed: 11557028
doi: 10.1016/S0005-2736(01)00384-4
Nakauchi, J. et al. Cloning and characterization of a human brain Na+-independent transporter for small neutral amino acids that transports D-serine with high affinity. Neurosci. Lett. 287, 231–235 (2000).
pubmed: 10863037
doi: 10.1016/S0304-3940(00)01169-1
Fukasawa, Y. et al. Identification and characterization of a Na(+)-independent neutral amino acid transporter that associates with the 4F2 heavy chain and exhibits substrate selectivity for small neutral D- and L-amino acids. J. Biol. Chem. 275, 9690–9698 (2000).
pubmed: 10734121
doi: 10.1074/jbc.275.13.9690
Ehmsen, J. T. et al. The astrocytic transporter SLC7A10 (Asc-1) mediates glycinergic inhibition of spinal cord motor neurons. Sci. Rep. 6, 35592 (2016).
pubmed: 27759100
pmcid: 5069678
doi: 10.1038/srep35592
Gauthier-Coles, G., Fairweather, S. J., Broer, A. & Broer, S. Do Amino Acid Antiporters Have Asymmetric Substrate Specificity? Biomolecules 13, 301 (2023).
pubmed: 36830670
pmcid: 9953452
doi: 10.3390/biom13020301
Rutter, A. R. et al. Evidence from gene knockout studies implicates Asc-1 as the primary transporter mediating d-serine reuptake in the mouse CNS. Eur. J. Neurosci. 25, 1757–1766 (2007).
pubmed: 17432963
doi: 10.1111/j.1460-9568.2007.05446.x
Safory, H. et al. The alanine-serine-cysteine-1 (Asc-1) transporter controls glycine levels in the brain and is required for glycinergic inhibitory transmission. EMBO Rep. 16, 590–598 (2015).
pubmed: 25755256
pmcid: 4428048
doi: 10.15252/embr.201439561
Xie, X. et al. Lack of the alanine-serine-cysteine transporter 1 causes tremors, seizures, and early postnatal death in mice. Brain Res. 1052, 212–221 (2005).
pubmed: 16026768
doi: 10.1016/j.brainres.2005.06.039
Hansen, K. B. et al. Structure, function, and pharmacology of glutamate receptor ion channels. Pharmacol. Rev. 73, 298–487 (2021).
pubmed: 34753794
pmcid: 8626789
doi: 10.1124/pharmrev.120.000131
Furukawa, H. & Gouaux, E. Mechanisms of activation, inhibition and specificity: crystal structures of the NMDA receptor NR1 ligand-binding core. EMBO J. 22, 2873–2885 (2003).
pubmed: 12805203
pmcid: 162155
doi: 10.1093/emboj/cdg303
Brown, J. M. et al. In vitro Characterization of a small molecule inhibitor of the alanine serine cysteine transporter -1 (SLC7A10). J. Neurochem. 129, 275–283 (2014).
pubmed: 24266811
doi: 10.1111/jnc.12618
Torrecillas, I. R. et al. Inhibition of the alanine-serine-cysteine-1 transporter by BMS-466442. ACS Chem. Neurosci. 10, 2510–2517 (2019).
pubmed: 30821959
doi: 10.1021/acschemneuro.9b00019
Mikou, A. et al. Asc-1 Transporter (SLC7A10): Homology Models And Molecular Dynamics Insights Into The First Steps Of The Transport Mechanism. Sci Rep 10, 3731 (2020).
pubmed: 32111919
pmcid: 7048771
doi: 10.1038/s41598-020-60617-y
Fujita, Y., Ishima, T. & Hashimoto, K. Supplementation with D-serine prevents the onset of cognitive deficits in adult offspring after maternal immune activation. Sci. Rep. 6, 37261 (2016).
pubmed: 27853241
pmcid: 5112512
doi: 10.1038/srep37261
Sakimura, K., Nakao, K., Yoshikawa, M., Suzuki, M. & Kimura, H. A novel Na(+) -Independent alanine-serine-cysteine transporter 1 inhibitor inhibits both influx and efflux of D-Serine. J. Neurosci. Res. 94, 888–895 (2016).
pubmed: 27302861
doi: 10.1002/jnr.23772
Labrie, V., Wong, A. H. & Roder, J. C. Contributions of the D-serine pathway to schizophrenia. Neuropharmacology 62, 1484–1503 (2012).
pubmed: 21295046
doi: 10.1016/j.neuropharm.2011.01.030
Nunes, E. A. et al. D-serine and schizophrenia: an update. Expert Rev. Neurother 12, 801–812 (2012).
pubmed: 22853788
doi: 10.1586/ern.12.65
Bendikov, I. et al. A CSF and postmortem brain study of D-serine metabolic parameters in schizophrenia. Schizophr. Res. 90, 41–51 (2007).
pubmed: 17156977
doi: 10.1016/j.schres.2006.10.010
Billard, J. M. & Freret, T. Asc-1 transporter activation: an alternative to rescue age-related alterations in functional plasticity at rat hippocampal CA3/CA1 synapses. J. Neurochem. 147, 514–525 (2018).
pubmed: 30187927
doi: 10.1111/jnc.14586
Jersin, R. A., Jonassen, L. R. & Dankel, S. N. The neutral amino acid transporter SLC7A10 in adipose tissue, obesity and insulin resistance. Front. Cell Dev. Biol. 10, 974338 (2022).
pubmed: 36172277
pmcid: 9512047
doi: 10.3389/fcell.2022.974338
Suwandhi, L. et al. Asc-1 regulates white versus beige adipocyte fate in a subcutaneous stromal cell population. Nat. Commun. 12, 1588 (2021).
pubmed: 33707431
pmcid: 7952576
doi: 10.1038/s41467-021-21826-9
Yan, R. et al. Mechanism of substrate transport and inhibition of the human LAT1-4F2hc amino acid transporter. Cell Discov. 7, 16 (2021).
pubmed: 33758168
pmcid: 7988154
doi: 10.1038/s41421-021-00247-4
Lee, Y. et al. Cryo-EM structure of the human L-type amino acid transporter 1 in complex with glycoprotein CD98hc. Nat. Struct. Mol. Biol. 26, 510–517 (2019).
pubmed: 31160781
doi: 10.1038/s41594-019-0237-7
Meury, M. et al. Detergent-induced stabilization and improved 3D map of the human heteromeric amino acid transporter 4F2hc-LAT2. PLoS One 9, e109882 (2014).
pubmed: 25299125
pmcid: 4192586
doi: 10.1371/journal.pone.0109882
Yan, R. H., Zhou, J. Y., Li, Y. N., Lei, J. L. & Zhou, Q. Structural insight into the substrate recognition and transport mechanism of the human LAT2-4F2hc complex. Cell Discov 6, 82 (2020).
pubmed: 33298890
pmcid: 7653941
doi: 10.1038/s41421-020-00207-4
Rosell, A. et al. Structural bases for the interaction and stabilization of the human amino acid transporter LAT2 with its ancillary protein 4F2hc. Proc. Natl. Acad. Sci. USA 111, 2966–2971 (2014).
pubmed: 24516142
pmcid: 3939895
doi: 10.1073/pnas.1323779111
Jeckelmann, J. M. & Fotiadis, D. Volta phase plate cryo-EM structure of the human heterodimeric amino acid transporter 4F2hc-LAT2. Int. J. Mol. Sci. 20, 931 (2019).
pubmed: 30795505
pmcid: 6413005
doi: 10.3390/ijms20040931
Yan, R. et al. Cryo-EM structure of the human heteromeric amino acid transporter b(0,+)AT-rBAT. Sci. Adv. 6, eaay6379 (2020).
pubmed: 32494597
pmcid: 7159911
doi: 10.1126/sciadv.aay6379
Wu, D. et al. Structural basis for amino acid exchange by a human heteromeric amino acid transporter. Proc. Natl. Acad. Sci. USA 117, 21281–21287 (2020).
pubmed: 32817565
pmcid: 7474693
doi: 10.1073/pnas.2008111117
Oda, K. et al. Consensus mutagenesis approach improves the thermal stability of system xc (-) transporter, xCT, and enables cryo-EM analyses. Protein Sci. 29, 2398–2407 (2020).
pubmed: 33016372
pmcid: 7679960
doi: 10.1002/pro.3966
Yan, R. et al. The structure of erastin-bound xCT-4F2hc complex reveals molecular mechanisms underlying erastin-induced ferroptosis. Cell Res. 32, 687–690 (2022).
pubmed: 35352032
pmcid: 9253326
doi: 10.1038/s41422-022-00642-w
Kazmier, K., Claxton, D. P. & McHaourab, H. S. Alternating access mechanisms of LeuT-fold transporters: trailblazing towards the promised energy landscapes. Current Opin. Struct. Biol. 45, 100–108 (2017).
doi: 10.1016/j.sbi.2016.12.006
Fang, Y. et al. Structure of a prokaryotic virtual proton pump at 3.2 A resolution. Nature 460, 1040–1043 (2009).
pubmed: 19578361
pmcid: 2745212
doi: 10.1038/nature08201
Gao, X. et al. Structure and mechanism of an amino acid antiporter. Science 324, 1565–1568 (2009).
pubmed: 19478139
doi: 10.1126/science.1173654
Gao, X. et al. Mechanism of substrate recognition and transport by an amino acid antiporter. Nature 463, 828–832 (2010).
pubmed: 20090677
doi: 10.1038/nature08741
Ma, D. et al. Structure and mechanism of a glutamate-GABA antiporter. Nature 483, 632–636 (2012).
pubmed: 22407317
doi: 10.1038/nature10917
Shaffer, P. L., Goehring, A., Shankaranarayanan, A. & Gouaux, E. Structure and mechanism of a Na+-independent amino acid transporter. Science 325, 1010–1014 (2009).
pubmed: 19608859
pmcid: 2851542
doi: 10.1126/science.1176088
Lei, J. & Frank, J. Automated acquisition of cryo-electron micrographs for single particle reconstruction on an FEI Tecnai electron microscope. J. Struct. Biol. 150, 69–80 (2005).
pubmed: 15797731
doi: 10.1016/j.jsb.2005.01.002
Zheng, S. Q. et al. MotionCor2: anisotropic correction of beam-induced motion for improved cryo-electron microscopy. Nature Methods 14, 331–332 (2017).
pubmed: 28250466
pmcid: 5494038
doi: 10.1038/nmeth.4193
Grant, T. & Grigorieff, N. Measuring the optimal exposure for single particle cryo-EM using a 2.6 Å reconstruction of rotavirus VP6. eLife 4, e06980 (2015).
pubmed: 26023829
pmcid: 4471936
doi: 10.7554/eLife.06980
Zhang, K. Gctf: real-time CTF determination and correction. J. Struct. Biol. 193, 1–12 (2016).
pubmed: 26592709
pmcid: 4711343
doi: 10.1016/j.jsb.2015.11.003
Punjani, A., Rubinstein, J. L., Fleet, D. J. & Brubaker, M. A. cryoSPARC: algorithms for rapid unsupervised cryo-EM structure determination. Nature Methods 14, 290–296 (2017).
pubmed: 28165473
doi: 10.1038/nmeth.4169
Rosenthal, P. B. & Henderson, R. Optimal determination of particle orientation, absolute hand, and contrast loss in single-particle electron cryomicroscopy. J. Mol. Biol. 333, 721–745 (2003).
pubmed: 14568533
doi: 10.1016/j.jmb.2003.07.013
Chen, S. et al. High-resolution noise substitution to measure overfitting and validate resolution in 3D structure determination by single particle electron cryomicroscopy. Ultramicroscopy 135, 24–35 (2013).
pubmed: 23872039
pmcid: 3834153
doi: 10.1016/j.ultramic.2013.06.004
Adams, P. D. et al. PHENIX: a comprehensive Python-based system for macromolecular structure solution. Acta Crystallogr. D Biol. Crystallogr. 66, 213–221 (2010).
pubmed: 20124702
pmcid: 2815670
doi: 10.1107/S0907444909052925
Emsley, P., Lohkamp, B., Scott, W. G. & Cowtan, K. Features and development of Coot. Acta Crystallogr. D Biol. Crystallogr. 66, 486–501 (2010).
pubmed: 20383002
pmcid: 2852313
doi: 10.1107/S0907444910007493
Trabuco, L. G., Villa, E., Mitra, K., Frank, J. & Schulten, K. Flexible fitting of atomic structures into electron microscopy maps using molecular dynamics. Structure 16, 673–683 (2008).
pubmed: 18462672
pmcid: 2430731
doi: 10.1016/j.str.2008.03.005
Broer, S. Xenopus laevis Oocytes. Methods Mol Biol 227, 245–258 (2003).
pubmed: 12824652