ATP modulates SLC7A5 (LAT1) synergistically with cholesterol.
Journal
Scientific reports
ISSN: 2045-2322
Titre abrégé: Sci Rep
Pays: England
ID NLM: 101563288
Informations de publication
Date de publication:
07 10 2020
07 10 2020
Historique:
received:
22
04
2020
accepted:
17
09
2020
entrez:
8
10
2020
pubmed:
9
10
2020
medline:
15
12
2020
Statut:
epublish
Résumé
The plasma membrane transporter hLAT1 is responsible for providing cells with essential amino acids. hLAT1 is over-expressed in virtually all human cancers making the protein a hot-spot in the fields of cancer and pharmacology research. However, regulatory aspects of hLAT1 biology are still poorly understood. A remarkable stimulation of transport activity was observed in the presence of physiological levels of cholesterol together with a selective increase of the affinity for the substrate on the internal site, suggesting a stabilization of the inward open conformation of hLAT1. A synergistic effect by ATP was also observed only in the presence of cholesterol. The same phenomenon was detected with the native protein. Altogether, the biochemical assays suggested that cholesterol and ATP binding sites are close to each other. The computational analysis identified two neighboring regions, one hydrophobic and one hydrophilic, to which cholesterol and ATP were docked, respectively. The computational data predicted interaction of the ϒ-phosphate of ATP with Lys 204, which was confirmed by site-directed mutagenesis. The hLAT1-K204Q mutant showed an impaired function and response to ATP. Interestingly, this residue is conserved in several members of the SLC7 family.
Identifiants
pubmed: 33028978
doi: 10.1038/s41598-020-73757-y
pii: 10.1038/s41598-020-73757-y
pmc: PMC7541457
doi:
Substances chimiques
Large Neutral Amino Acid-Transporter 1
0
Liposomes
0
SLC7A5 protein, human
0
Adenosine Triphosphate
8L70Q75FXE
Cholesterol
97C5T2UQ7J
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
16738Références
Fotiadis, D., Kanai, Y. & Palacin, M. The SLC3 and SLC7 families of amino acid transporters. Mol. Aspects Med. 34, 139–158. https://doi.org/10.1016/j.mam.2012.10.007 (2013).
doi: 10.1016/j.mam.2012.10.007
pubmed: 23506863
Scalise, M., Galluccio, M., Console, L., Pochini, L. & Indiveri, C. The Human SLC7A5 (LAT1): The intriguing histidine/large neutral amino acid transporter and its relevance to human health. Front. Chem. 6, 243. https://doi.org/10.3389/fchem.2018.00243 (2018).
doi: 10.3389/fchem.2018.00243
pubmed: 29988369
pmcid: 6023973
Napolitano, L. et al. LAT1 is the transport competent unit of the LAT1/CD98 heterodimeric amino acid transporter. Int. J. Biochem. Cell Biol. 67, 25–33. https://doi.org/10.1016/j.biocel.2015.08.004 (2015).
doi: 10.1016/j.biocel.2015.08.004
pubmed: 26256001
Cantor, J. M. & Ginsberg, M. H. CD98 at the crossroads of adaptive immunity and cancer. J. Cell Sci. 125, 1373–1382. https://doi.org/10.1242/jcs.096040 (2012).
doi: 10.1242/jcs.096040
pubmed: 22499670
pmcid: 3336374
Tarlungeanu, D. C. et al. Impaired amino acid transport at the blood brain barrier is a cause of autism spectrum disorder. Cell 167, 1481–1494.e1418. https://doi.org/10.1016/j.cell.2016.11.013 (2016).
doi: 10.1016/j.cell.2016.11.013
pubmed: 27912058
pmcid: 5554935
Ohgaki, R. et al. Essential roles of L-type amino acid transporter 1 in syncytiotrophoblast development by presenting fusogenic 4F2hc. Mol. Cell. Biol. https://doi.org/10.1128/MCB.00427-16 (2017).
doi: 10.1128/MCB.00427-16
pubmed: 28320871
pmcid: 5440655
Bhutia, Y. D., Babu, E., Ramachandran, S. & Ganapathy, V. Amino Acid transporters in cancer and their relevance to “glutamine addiction”: Novel targets for the design of a new class of anticancer drugs. Can. Res. 75, 1782–1788. https://doi.org/10.1158/0008-5472.CAN-14-3745 (2015).
doi: 10.1158/0008-5472.CAN-14-3745
Nicklin, P. et al. Bidirectional transport of amino acids regulates mTOR and autophagy. Cell 136, 521–534. https://doi.org/10.1016/j.cell.2008.11.044 (2009).
doi: 10.1016/j.cell.2008.11.044
pubmed: 3733119
pmcid: 3733119
Scalise, M., Pochini, L., Galluccio, M., Console, L. & Indiveri, C. Glutamine transport and mitochondrial metabolism in cancer cell growth. Front. Oncol. 7, 306. https://doi.org/10.3389/fonc.2017.00306 (2017).
doi: 10.3389/fonc.2017.00306
pubmed: 29376023
pmcid: 5770653
Broer, S. & Broer, A. Amino acid homeostasis and signalling in mammalian cells and organisms. Biochem. J. 474, 1935–1963. https://doi.org/10.1042/BCJ20160822 (2017).
doi: 10.1042/BCJ20160822
pubmed: 5444488
pmcid: 5444488
Wolfson, R. L. & Sabatini, D. M. The dawn of the age of amino acid sensors for the mTORC1 pathway. Cell Metab. 26, 301–309. https://doi.org/10.1016/j.cmet.2017.07.001 (2017).
doi: 10.1016/j.cmet.2017.07.001
pubmed: 28768171
pmcid: 5560103
Singh, N. & Ecker, G. F. Insights into the structure, function, and ligand discovery of the large neutral amino acid transporter 1, LAT1. Int. J. Mol. Sci. https://doi.org/10.3390/ijms19051278 (2018).
doi: 10.3390/ijms19051278
pubmed: 30577601
pmcid: 6337383
Singh, N. et al. Discovery of potent inhibitors for the large neutral amino acid transporter 1 (LAT1) by structure-based methods. Int. J. Mol. Sci. https://doi.org/10.3390/ijms20010027 (2018).
doi: 10.3390/ijms20010027
pubmed: 30577601
pmcid: 6337383
Singh, N., Villoutreix, B. O. & Ecker, G. F. Rigorous sampling of docking poses unveils binding hypothesis for the halogenated ligands of L-type Amino acid Transporter 1 (LAT1). Sci. Rep. 9, 15061. https://doi.org/10.1038/s41598-019-51455-8 (2019).
doi: 10.1038/s41598-019-51455-8
pubmed: 31636293
pmcid: 6803698
Yan, R., Zhao, X., Lei, J. & Zhou, Q. Structure of the human LAT1-4F2hc heteromeric amino acid transporter complex. Nature 568, 127–130. https://doi.org/10.1038/s41586-019-1011-z (2019).
doi: 10.1038/s41586-019-1011-z
pubmed: 30867591
Oda, K. et al. L-type amino acid transporter 1 inhibitors inhibit tumor cell growth. Cancer Sci. 101, 173–179. https://doi.org/10.1111/j.1349-7006.2009.01386.x (2010).
doi: 10.1111/j.1349-7006.2009.01386.x
pubmed: 19900191
Okano, N. et al. First-in-human phase I study of JPH203, an L-type amino acid transporter 1 inhibitor, in patients with advanced solid tumors. Invest. New Drugs https://doi.org/10.1007/s10637-020-00924-3 (2020).
doi: 10.1007/s10637-020-00924-3
pubmed: 32198649
del Amo, E. M., Urtti, A. & Yliperttula, M. Pharmacokinetic role of L-type amino acid transporters LAT1 and LAT2. Eur. J. Pharm. Sci. Off. J. Eur. Federation Pharm. Sci. 35, 161–174. https://doi.org/10.1016/j.ejps.2008.06.015 (2008).
doi: 10.1016/j.ejps.2008.06.015
Scalise, M., Pochini, L., Giangregorio, N., Tonazzi, A. & Indiveri, C. Proteoliposomes as tool for assaying membrane transporter functions and interactions with xenobiotics. Pharmaceutics 5, 472–497. https://doi.org/10.3390/pharmaceutics5030472 (2013).
doi: 10.3390/pharmaceutics5030472
pubmed: 24300519
pmcid: 3836619
Dickens, D. et al. Modulation of LAT1 (SLC7A5) transporter activity and stability by membrane cholesterol. Sci. Rep. 7, 43580. https://doi.org/10.1038/srep43580 (2017).
doi: 10.1038/srep43580
pubmed: 28272458
pmcid: 5341093
Napolitano, L. et al. Novel insights into the transport mechanism of the human amino acid transporter LAT1 (SLC7A5). Probing critical residues for substrate translocation. Biochim. Biophys. Acta General Subjects 1861, 727–736. https://doi.org/10.1016/j.bbagen.2017.01.013 (2017).
doi: 10.1016/j.bbagen.2017.01.013
pubmed: 28088504
Cosco, J., Regina, T. M. R., Scalise, M., Galluccio, M. & Indiveri, C. Regulatory aspects of the vacuolar CAT2 arginine transporter of S. lycopersicum: Role of osmotic pressure and cations. Int. J. Mol. Sci. https://doi.org/10.3390/ijms20040906 (2019).
doi: 10.3390/ijms20040906
pubmed: 30791488
pmcid: 6413183
Pochini, L. et al. The human OCTN1 (SLC22A4) reconstituted in liposomes catalyzes acetylcholine transport which is defective in the mutant L503F associated to the Crohn’s disease. Biochem. Biophys. Acta. 559–565, 2012. https://doi.org/10.1016/j.bbamem.2011.12.014 (1818).
doi: 10.1016/j.bbamem.2011.12.014
Levine, K. B., Cloherty, E. K., Hamill, S. & Carruthers, A. Molecular determinants of sugar transport regulation by ATP. Biochemistry 41, 12629–12638. https://doi.org/10.1021/bi0258997 (2002).
doi: 10.1021/bi0258997
pubmed: 12379105
Echtay, K. S. et al. Uncoupling proteins: Martin Klingenberg’s contributions for 40 years. Arch. Biochem. Biophys. 657, 41–55. https://doi.org/10.1016/j.abb.2018.09.006 (2018).
doi: 10.1016/j.abb.2018.09.006
pubmed: 30217511
Fitz, J. G. Regulation of cellular ATP release. Trans. Am. Clin. Climatol. Assoc. 118, 199–208 (2007).
pubmed: 18528503
pmcid: 1863605
Traut, T. W. Physiological concentrations of purines and pyrimidines. Mol. Cell. Biochem. 140, 1–22. https://doi.org/10.1007/bf00928361 (1994).
doi: 10.1007/bf00928361
pubmed: 7877593
Falzoni, S., Donvito, G. & Di Virgilio, F. Detecting adenosine triphosphate in the pericellular space. Interface Focus 3, 20120101. https://doi.org/10.1098/rsfs.2012.0101 (2013).
doi: 10.1098/rsfs.2012.0101
pubmed: 23853707
pmcid: 3638417
Halgren, T. New method for fast and accurate binding-site identification and analysis. Chem. Biol. Drug Des. 69, 146–148. https://doi.org/10.1111/j.1747-0285.2007.00483.x (2007).
doi: 10.1111/j.1747-0285.2007.00483.x
pubmed: 17381729
Halgren, T. A. Identifying and characterizing binding sites and assessing druggability. J. Chem. Inf. Model. 49, 377–389. https://doi.org/10.1021/ci800324m (2009).
doi: 10.1021/ci800324m
pubmed: 19434839
Fantini, J. & Barrantes, F. J. How cholesterol interacts with membrane proteins: an exploration of cholesterol-binding sites including CRAC, CARC, and tilted domains. Front. Physiol. 4, 31. https://doi.org/10.3389/fphys.2013.00031 (2013).
doi: 10.3389/fphys.2013.00031
pubmed: 23450735
pmcid: 3584320
Trott, O. & Olson, A. J. AutoDock Vina: improving the speed and accuracy of docking with a new scoring function, efficient optimization, and multithreading. J. Comput. Chem. 31, 455–461. https://doi.org/10.1002/jcc.21334 (2010).
doi: 10.1002/jcc.21334
pubmed: 19499576
pmcid: 3041641
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. https://doi.org/10.1074/jbc.273.37.23629 (1998).
doi: 10.1074/jbc.273.37.23629
pubmed: 9726963
Penmatsa, A., Wang, K. H. & Gouaux, E. X-ray structure of dopamine transporter elucidates antidepressant mechanism. Nature 503, 85–90. https://doi.org/10.1038/nature12533 (2013).
doi: 10.1038/nature12533
pubmed: 24037379
pmcid: 3904663
Laursen, L. et al. Cholesterol binding to a conserved site modulates the conformation, pharmacology, and transport kinetics of the human serotonin transporter. J. Biol. Chem. 293, 3510–3523. https://doi.org/10.1074/jbc.M117.809046 (2018).
doi: 10.1074/jbc.M117.809046
pubmed: 29352106
pmcid: 5846164
Coleman, J. A., Green, E. M. & Gouaux, E. X-ray structures and mechanism of the human serotonin transporter. Nature 532, 334–339. https://doi.org/10.1038/nature17629 (2016).
doi: 10.1038/nature17629
pubmed: 27049939
pmcid: 4898786
Pochini, L. et al. Effect of cholesterol on the organic cation transporter OCTN1 (SLC22A4). Int. J. Mol. Sci. https://doi.org/10.3390/ijms21031091 (2020).
doi: 10.3390/ijms21031091
pubmed: 32041338
pmcid: 7037232
Scalise, M. et al. Insights into the transport side of the human SLC38A9 transceptor. Biochim. Biophys. Acta 1558–1567, 2019. https://doi.org/10.1016/j.bbamem.2019.07.006 (1861).
doi: 10.1016/j.bbamem.2019.07.006
Scalise, M. et al. Interaction of cholesterol with the human SLC1A5 (ASCT2): Insights into structure/function relationships. Front. Mol. Biosci. 6, 110. https://doi.org/10.3389/fmolb.2019.00110 (2019).
doi: 10.3389/fmolb.2019.00110
pubmed: 31709262
pmcid: 6819821
Castellano, B. M. et al. Lysosomal cholesterol activates mTORC1 via an SLC38A9-Niemann-Pick C1 signaling complex. Science 355, 1306–1311. https://doi.org/10.1126/science.aag1417 (2017).
doi: 10.1126/science.aag1417
pubmed: 28336668
pmcid: 5823611
Hormann, S., Gai, Z., Kullak-Ublick, G. A. & Visentin, M. Plasma membrane cholesterol regulates the allosteric binding of 1-methyl-4-phenylpyridinium to organic cation transporter 2 (SLC22A2). J. Pharmacol. Exp. Therap. 372, 46–53. https://doi.org/10.1124/jpet.119.260877 (2020).
doi: 10.1124/jpet.119.260877
Barnes, K., Ingram, J. C., Bennett, M. D., Stewart, G. W. & Baldwin, S. A. Methyl-beta-cyclodextrin stimulates glucose uptake in Clone 9 cells: a possible role for lipid rafts. Biochem. J. 378, 343–351. https://doi.org/10.1042/BJ20031186 (2004).
doi: 10.1042/BJ20031186
pubmed: 14616090
pmcid: 1223971
Zeppelin, T., Ladefoged, L. K., Sinning, S., Periole, X. & Schiott, B. A direct interaction of cholesterol with the dopamine transporter prevents its out-to-inward transition. PLoS Comput. Biol. 14, e1005907. https://doi.org/10.1371/journal.pcbi.1005907 (2018).
doi: 10.1371/journal.pcbi.1005907
pubmed: 29329285
pmcid: 5811071
Garcia, A. et al. Cholesterol depletion inhibits Na(+), K(+)-ATPase activity in a near-native membrane environment. J. Biol. Chem. 294, 5956–5969. https://doi.org/10.1074/jbc.RA118.006223 (2019).
doi: 10.1074/jbc.RA118.006223
pubmed: 30770471
pmcid: 6463725
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. https://doi.org/10.1038/s41594-019-0237-7 (2019).
doi: 10.1038/s41594-019-0237-7
pubmed: 31160781
Kaczmarski, J. A. et al. Structural basis for the allosteric regulation of the sbta bicarbonate transporter by the PII-like protein, SbtB, from Cyanobium sp. PCC7001. Biochemistry 58, 5030–5039. https://doi.org/10.1021/acs.biochem.9b00880 (2019).
doi: 10.1021/acs.biochem.9b00880
pubmed: 31746199
Bertholet, A. M. & Kirichok, Y. UCP1: A transporter for H(+) and fatty acid anions. Biochimie 134, 28–34. https://doi.org/10.1016/j.biochi.2016.10.013 (2017).
doi: 10.1016/j.biochi.2016.10.013
pubmed: 27984203
Errasti-Murugarren, E. et al. L amino acid transporter structure and molecular bases for the asymmetry of substrate interaction. Nat. Commun. 10, 1807. https://doi.org/10.1038/s41467-019-09837-z (2019).
doi: 10.1038/s41467-019-09837-z
pubmed: 31000719
pmcid: 6472337
Shaffer, P. L., Goehring, A., Shankaranarayanan, A. & Gouaux, E. Structure and mechanism of a Na+-independent amino acid transporter. Science 325, 1010–1014. https://doi.org/10.1126/science.1176088 (2009).
doi: 10.1126/science.1176088
pubmed: 19608859
pmcid: 2851542
Jungnickel, K. E. J., Parker, J. L. & Newstead, S. Structural basis for amino acid transport by the CAT family of SLC7 transporters. Nat. Commun. 9, 550. https://doi.org/10.1038/s41467-018-03066-6 (2018).
doi: 10.1038/s41467-018-03066-6
pubmed: 29416041
pmcid: 5803215
Palazzolo, L. et al. In silico description of LAT1 transport mechanism at an atomistic level. Front. Chem. 6, 350. https://doi.org/10.3389/fchem.2018.00350 (2018).
doi: 10.3389/fchem.2018.00350
pubmed: 30197880
pmcid: 6117385
Cantor, J. R. & Sabatini, D. M. Cancer cell metabolism: One hallmark, many faces. Cancer Discov. 2, 881–898. https://doi.org/10.1158/2159-8290.CD-12-0345 (2012).
doi: 10.1158/2159-8290.CD-12-0345
pubmed: 23009760
pmcid: 3491070
Ho, S. N., Hunt, H. D., Horton, R. M., Pullen, J. K. & Pease, L. R. Site-directed mutagenesis by overlap extension using the polymerase chain reaction. Gene 77, 51–59. https://doi.org/10.1016/0378-1119(89)90358-2 (1989).
doi: 10.1016/0378-1119(89)90358-2
pubmed: 2744487
Galluccio, M. et al. Functional and molecular effects of mercury compounds on the human OCTN1 cation transporter: C50 and C136 are the targets for potent inhibition. Toxicol. Sci. Off. J. Society Toxicol. 144, 105–113. https://doi.org/10.1093/toxsci/kfu259 (2015).
doi: 10.1093/toxsci/kfu259
Galluccio, M., Pingitore, P., Scalise, M. & Indiveri, C. Cloning, large scale over-expression in E. coli and purification of the components of the human LAT 1 (SLC7A5) amino acid transporter. Protein J. 32, 442–448. https://doi.org/10.1007/s10930-013-9503-4 (2013).
doi: 10.1007/s10930-013-9503-4
pubmed: 23912240
Massey, J. B. Effect of cholesteryl hemisuccinate on the interfacial properties of phosphatidylcholine bilayers. Biochem. Biophys. Acta. 1415, 193–204. https://doi.org/10.1016/s0005-2736(98)00194-1 (1998).
doi: 10.1016/s0005-2736(98)00194-1
pubmed: 9858729
Indiveri, C., Prezioso, G., Dierks, T., Kramer, R. & Palmieri, F. Kinetic characterization of the reconstituted dicarboxylate carrier from mitochondria: A four-binding-site sequential transport system. Biochem. Biophys. Acta. 1143, 310–318. https://doi.org/10.1016/0005-2728(93)90202-q (1993).
doi: 10.1016/0005-2728(93)90202-q
pubmed: 8329439
Schrödinger, R. Maestro, Schrödinger, LLC, New York, NY (2019).
Schrödinger, R. Protein Preparation Wizard; Epik, Schrödinger, LLC, New York, NY, 2016; Impact, Schrödinger, LLC, New York, NY, 2016; Prime, Schrödinger, LLC, New York, NY (2019).
Sastry, G. M., Adzhigirey, M., Day, T., Annabhimoju, R. & Sherman, W. Protein and ligand preparation: Parameters, protocols, and influence on virtual screening enrichments. J. Comput. Aided Mol. Des. 27, 221–234. https://doi.org/10.1007/s10822-013-9644-8 (2013).
doi: 10.1007/s10822-013-9644-8
pubmed: 23579614
Schrödinger, R SiteMap, Schrödinger, LLC, New York, NY (2019).
Schrödinger, R. LigPrep, Schrödinger, LLC, New York, NY (2019).
Halgren, T. A. et al. Glide: A new approach for rapid, accurate docking and scoring. 2. Enrichment factors in database screening. J. Med. Chem. 47, 1750–1759. https://doi.org/10.1021/jm030644s (2004).
doi: 10.1021/jm030644s
pubmed: 15027866
Shelley, J. C. et al. Epik: A software program for pK(a) prediction and protonation state generation for drug-like molecules. J. Comput. Aided Mol. Des. 21, 681–691. https://doi.org/10.1007/s10822-007-9133-z (2007).
doi: 10.1007/s10822-007-9133-z
pubmed: 17899391
Schrödinger, R. Induced Fit Docking protocol; Glide, Schrödinger, LLC, New York, NY, 2016; Prime, Schrödinger, LLC, New York, NY (2019).
Farid, R., Day, T., Friesner, R. A. & Pearlstein, R. A. New insights about HERG blockade obtained from protein modeling, potential energy mapping, and docking studies. Bioorg. Med. Chem. 14, 3160–3173. https://doi.org/10.1016/j.bmc.2005.12.032 (2006).
doi: 10.1016/j.bmc.2005.12.032
pubmed: 16413785
Sherman, W., Day, T., Jacobson, M. P., Friesner, R. A. & Farid, R. Novel procedure for modeling ligand/receptor induced fit effects. J. Med. Chem. 49, 534–553. https://doi.org/10.1021/jm050540c (2006).
doi: 10.1021/jm050540c
pubmed: 16420040
Friesner, R. A. et al. Glide: A new approach for rapid, accurate docking and scoring. 1. Method and assessment of docking accuracy. J. Med. Chem. 47, 1739–1749. https://doi.org/10.1021/jm0306430 (2004).
doi: 10.1021/jm0306430
pubmed: 15027865
Genheden, S. & Ryde, U. The MM/PBSA and MM/GBSA methods to estimate ligand-binding affinities. Expert Opin. Drug Discov. 10, 449–461. https://doi.org/10.1517/17460441.2015.1032936 (2015).
doi: 10.1517/17460441.2015.1032936
pubmed: 25835573
pmcid: 4487606
Li, J. et al. The VSGB 2.0 model: A next generation energy model for high resolution protein structure modeling. Proteins 79, 2794–2812. https://doi.org/10.1002/prot.23106 (2011).
doi: 10.1002/prot.23106
pubmed: 21905107
pmcid: 3206729
Pettersen, E. F. et al. UCSF Chimera—A visualization system for exploratory research and analysis. J. Comput. Chem. 25, 1605–1612. https://doi.org/10.1002/jcc.20084 (2004).
doi: 10.1002/jcc.20084
Madeira, F. et al. The EMBL-EBI search and sequence analysis tools APIs in 2019. Nucleic Acids Res. 47, W636–W641. https://doi.org/10.1093/nar/gkz268 (2019).
doi: 10.1093/nar/gkz268
pubmed: 6602479
pmcid: 6602479
Chien, H. C. et al. Reevaluating the substrate specificity of the L-type amino acid transporter (LAT1). J. Med. Chem. 61, 7358–7373. https://doi.org/10.1021/acs.jmedchem.8b01007 (2018).
doi: 10.1021/acs.jmedchem.8b01007
pubmed: 30048132
pmcid: 6668346
Sali, A. & Blundell, T. L. Comparative protein modelling by satisfaction of spatial restraints. J. Mol. Biol. 234, 779–815. https://doi.org/10.1006/jmbi.1993.1626 (1993).
doi: 10.1006/jmbi.1993.1626
pubmed: 8254673
Ilgu, H. et al. Insights into the molecular basis for substrate binding and specificity of the wild-type L-arginine/agmatine antiporter AdiC. Proc. Natl. Acad. Sci. USA 113, 10358–10363. https://doi.org/10.1073/pnas.1605442113 (2016).
doi: 10.1073/pnas.1605442113
pubmed: 27582465