Substrate discrimination by ergothioneine transporter SLC22A4 and carnitine transporter SLC22A5: Gain-of-function by interchange of selected amino acids
Open accessAbstract
ETT (originally designated as OCTN1; human gene symbol SLC22A4) and CTT (OCTN2; SLC22A5) are highly specific transporters of ergothioneine and carnitine, respectively. Despite a high degree of sequence homology, both carriers discriminate precisely between substrates: ETT does not transport carnitine, and CTT does not transport ergothioneine. Our aim was to turn ETT into a transporter for carnitine and CTT into a transporter for ergothioneine by a limited number of point mutations. From a multiple alignment of several mammalian amino acid sequences, those positions were selected for conversion that were momentously different between ETT and CTT from human but conserved among all orthologues. Mutants were expressed in 293 cells and assayed for transport of ergothioneine and carnitine. Several ETT mutants clearly catalyzed transport of carnitine, up to 35% relative to wild-type CTT. Amazingly, complementary substitutions in CTT did not provoke transport activity for ergothioneine. In similar contrast, carnitine transport by CTT mutants was abolished by very few substitutions, whereas ergothioneine transport by ETT mutants was maintained even with the construct most active in carnitine transport. To explain these results, we propose that ETT and CTT use dissimilar pathways for conformational change, in addition to incongruent substrate binding sites. In other words, carnitine is excluded from ETT by binding, and ergothioneine is excluded from CTT by turnover movement. Our data indicate amino acids critical for substrate discrimination not only in transmembrane segments 5, 7, 8, and 10, but also in segments 9 and 12 which were hitherto considered as unimportant.
The claims and statistics of this paper haven’t been checked yet.
Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.
Integrity signals
Registry
Cites
Statistics
Claims the paper makes
The claims and statistics of this paper haven’t been checked yet.
Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.
Claims other papers make about this one
Each is checked against this paper’s own text: does it say what the citing paper says it does?
SLC22A4 and its homologues have significant specificity for ERG and weak activity for various drugs.
“Subsequent work(87,169–172) has confirmed and reinforced this view of SLC22A4 and its homologues(173) as having significant specificity for ERG, and weak activity for various drugs(174–177).”
From The biology of ergothioneine, an antioxidant nutraceutical (2020) · cites this paper as [169]
Self-check
Percentages, sums and reported differences re-computed from the paper’s own numbers.
The self-check needs the paper’s full text.
Cites
Structure and Mechanism of the Lactose Permease of Escherichia coli
Abramson, J., Smirnova, I., Kasho, V. N., Verner, G., Kaback, H. R., & Iwata, S. (2003) · Science · doi:10.1126/science.1088196 · Cited by 1,389 · see citing works
Structure and Mechanism of the Glycerol-3-Phosphate Transporter from Escherichia coli
Huang, Y., Lemieux, M. J., Song, J., Auer, M., & Wang, D. (2003) · Science · doi:10.1126/science.1087619 · Cited by 1,015 · see citing works
Discovery of the ergothioneine transporter
Gründemann, D., Harlfinger, S., Gölz, S., Geerts, A., Lazar, A., Berkels, R., Jung, N., RUBBERT, A., & Schömig, E. (2005) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.0408624102 · Cited by 613 · see citing works
Self-citationSelective substrates for non-neuronal monoamine transporters.
Gründemann, D., Liebich, G., Kiefer, N., Köster, S., & Schömig, E. (1999) · Molecular Pharmacology · doi:10.1124/mol.56.1.1 · Cited by 266 · see citing works
Self-citationGorboulev, V., Shatskaya, N., Volk, C., & Koepsell, H. (2005) · Molecular Pharmacology · doi:10.1124/mol.104.008821 · Cited by 105 · see citing works
Schömig, E., Lazar, A., & Gründemann, D. (2006) · Handbook of Experimental Pharmacology · doi:10.1007/3-540-29784-7_8 · Cited by 97 · see citing works
Self-citationLu, K., Nishimori, H., Nakamura, Y., Shima, K., & Kuwajima, M. (1998) · Biochemical and Biophysical Research Communications · doi:10.1006/bbrc.1998.9708 · Cited by 86 · see citing works
Seth, P., Wu, X., Huang, W. Y., Leibach, F. H., & Ganapathy, V. (1999) · Journal of Biological Chemistry · doi:10.1074/jbc.274.47.33388 · Cited by 85 · see citing works
Grigat, S., Fork, C., Bach, M., Gölz, S., Geerts, A., Schömig, E., & Gründemann, D. (2008) · Drug Metabolism and Disposition · doi:10.1124/dmd.108.023929 · Cited by 81 · see citing works
Self-citationMueckler, M. M., & Makepeace, C. M. (2009) · Biochemistry · doi:10.1021/bi900521n · Cited by 80 · see citing works
A Three-dimensional Model of Human Organic Anion Transporter 1
Perry, J. L., Dembla-Rajpal, N., Hall, L. A., & Pritchard, J. B. (2006) · Journal of Biological Chemistry · doi:10.1074/jbc.m608834200 · Cited by 80 · see citing works
Zhang, X., Shirahatti, N. V., Mahadevan, D., & Wright, S. H. (2005) · Journal of Biological Chemistry · doi:10.1074/jbc.m506342200 · Cited by 73 · see citing works
Grigat, S., Harlfinger, S., Pal, S., Striebinger, R., Golz, S., Geerts, A., Lazar, A., Schömig, E., & Gründemann, D. (2007) · Biochemical Pharmacology · doi:10.1016/j.bcp.2007.04.015 · Cited by 72 · see citing works
Self-citationFeng, B., Dresser, M. J., Shu, Y., Johns, S. J., & Giacomini, K. M. (2001) · Biochemistry · doi:10.1021/bi002841o · Cited by 72 · see citing works
Mueckler, M. M., & Makepeace, C. M. (2004) · Journal of Biological Chemistry · doi:10.1074/jbc.m310786200 · Cited by 69 · see citing works
Ohashi, R., Tamai, I., Inano, A., Katsura, M., Sai, Y., Nezu, J., & Tsuji, A. (2002) · Journal of Pharmacology and Experimental Therapeutics · doi:10.1124/jpet.102.036004 · Cited by 67 · see citing works
Structure and Transport Mechanism of the Bacterial Oxalate Transporter OxlT
Hirai, T., & Subramaniam, S. (2004) · Biophysical Journal · doi:10.1529/biophysj.104.049320 · Cited by 58 · see citing works
Cysteine-scanning Mutagenesis of Transmembrane Segment 7 of the GLUT1 Glucose Transporter
Hruz, P. W., & Mueckler, M. M. (1999) · Journal of Biological Chemistry · doi:10.1074/jbc.274.51.36176 · Cited by 58 · see citing works
1,1?-Diethyl-2,2?-cyanine (decynium22) potently inhibits the renal transport of organic cations
Sch�mig, E., Babin–Ebell, J., & Russ, H. (1993) · Naunyn-Schmiedeberg s Archives of Pharmacology · doi:10.1007/bf00165387 · Cited by 54 · see citing works
Self-citationEvolutionary processes and evolutionary noise at the molecular level
Zuckerkandl, E. (1976) · Journal of Molecular Evolution · doi:10.1007/bf01743626 · Cited by 51 · see citing works
Bach, M., Grigat, S., Pawlik, B., Fork, C., Utermöhlen, O., Pal, S., Banczyk, D., Lazar, A., Schömig, E., & Gründemann, D. (2007) · FEBS Journal · doi:10.1111/j.1742-4658.2006.05623.x · Cited by 49 · see citing works
Self-citationMueckler, M. M., & Makepeace, C. M. (2002) · Journal of Biological Chemistry · doi:10.1074/jbc.m109157200 · Cited by 48 · see citing works
Mueckler, M. M., & Makepeace, C. M. (1999) · Journal of Biological Chemistry · doi:10.1074/jbc.274.16.10923 · Cited by 48 · see citing works
Mueckler, M. M., & Makepeace, C. M. (2006) · Journal of Biological Chemistry · doi:10.1074/jbc.m608158200 · Cited by 37 · see citing works
Functional Domains in the Carnitine Transporter OCTN2, Defective in Primary Carnitine Deficiency
Filippo, C. A. D. S., Wang, Y., & Longo, N. (2003) · Journal of Biological Chemistry · doi:10.1074/jbc.m307911200 · Cited by 33 · see citing works
Feng, B., Shu, Y., & Giacomini, K. M. (2002) · Biochemistry · doi:10.1021/bi0200615 · Cited by 33 · see citing works
Hong, M., Zhou, F., Lee, K. C., & You, G. (2006) · Journal of Pharmacology and Experimental Therapeutics · doi:10.1124/jpet.106.117663 · Cited by 26 · see citing works
Inano, A., Sai, Y., Kato, Y., Tamai, I., Ishiguro, M., & Tsuji, A. (2004) · Drug Metabolism and Pharmacokinetics · doi:10.2133/dmpk.19.180 · Cited by 23 · see citing works
Rahbeeni, Z. A., Vaz, F. M., Al‐Hussein, K. A., Bucknall, M. P., Ruiter, J. P. N., Wanders, R. J. A., & Rashed, M. S. (2002) · Journal of Inherited Metabolic Disease · doi:10.1023/a:1020143632011 · Cited by 22 · see citing works
Flagged first, then by citations
Cited by
Sorted by citations
Organic Cation Transporters in Health and Disease
Koepsell, H. (2019) · Pharmacological Reviews · doi:10.1124/pr.118.015578 · Cited by 293 · see citing works
The biology of ergothioneine, an antioxidant nutraceutical
Borodina, I., Kenny, L. C., McCarthy, C. M., Paramasivan, K., Pretorius, E., Roberts, T. J., van der Hoek, S. A., & Kell, D. B. (2020) · Nutrition Research Reviews · doi:10.1017/s0954422419000301 · Cited by 296 · see citing works
“Subsequent work (87,169 – 172) has confirmed and reinforced this view of SLC22A4 and its homologues (173) as having significant specificity for ERG, and weak activity for various drugs (174 – 177) .”
BackgroundGlutathione analogs in prokaryotes.
Fahey, R. C. (2013) · Biochimica et Biophysica Acta · doi:10.1016/j.bbagen.2012.10.006 · Cited by 205 · see citing works
Cyanobacteria produce high levels of ergothioneine
Pfeiffer, C., Bauer, T., Surek, B., Schömig, E., & Gründemann, D. (2011) · Food Chemistry · doi:10.1016/j.foodchem.2011.06.047 · Cited by 153 · see citing works
Self-citationErgothioneine, an adaptive antioxidant for the protection of injured tissues? A hypothesis
Halliwell, B., Cheah, I. K., & Drum, C. L. (2016) · Biochemical and Biophysical Research Communications · doi:10.1016/j.bbrc.2015.12.124 · Cited by 123 · see citing works
The ergothioneine transporter controls and indicates ergothioneine activity — A review
Gründemann, D. (2012) · Preventive Medicine · doi:10.1016/j.ypmed.2011.12.001 · Cited by 111 · see citing works
Self-citationThe ergothioneine transporter (ETT): substrates and locations, an inventory
Gründemann, D., Hartmann, L., & Flögel, S. (2021) · FEBS Letters · doi:10.1002/1873-3468.14269 · Cited by 81 · see citing works
Self-citationOAT2 catalyses efflux of glutamate and uptake of orotic acid
Fork, C., Bauer, T., Gölz, S., Geerts, A., Weiland, J., Turco, D. D., Schömig, E., & Gründemann, D. (2011) · Biochemical Journal · doi:10.1042/bj20101904 · Cited by 69 · see citing works
Self-citationSLC22, SLC44, and SLC47 Transporters—Organic Anion and Cation Transporters
Pelis, R. M., & Wright, S. H. (2014) · Current topics in membranes · doi:10.1016/b978-0-12-800223-0.00006-2 · Cited by 68 · see citing works
Substrate Selectivity Check of the Ergothioneine Transporter
Tschirka, J., Kreisor, M., Betz, J., & Gründemann, D. (2018) · Drug Metabolism and Disposition · doi:10.1124/dmd.118.080440 · Cited by 66 · see citing works
Self-citationNemkov, T. G., Key, A., Stephenson, D., Earley, E. J., Keele, G. R., Hay, A. M., Amireault, P., Casimir, M., Dussiot, M., Dzieciątkowska, M., et al. (2024) · Blood · doi:10.1182/blood.2024023983 · Cited by 55 · see citing works
Ishimoto, T., Nakamichi, N., Hosotani, H., Masuo, Y., Sugiura, T., & Kato, Y. (2014) · PLoS ONE · doi:10.1371/journal.pone.0089434 · Cited by 55 · see citing works
Stoffels, C., Oumari, M., Perrou, A., Termath, A. O., Schlundt, W., Schmalz, H., Schäfer, M., Wewer, V., Metzger, S. U., Schömig, E., et al. (2017) · Free Radical Biology and Medicine · doi:10.1016/j.freeradbiomed.2017.10.372 · Cited by 54 · see citing works
Self-citationKnockout of the ergothioneine transporter ETT in zebrafish results in increased 8-oxoguanine levels
Pfeiffer, C., Bach, M., Bauer, T., da Ponte, J. C., Schömig, E., & Gründemann, D. (2015) · Free Radical Biology and Medicine · doi:10.1016/j.freeradbiomed.2015.02.026 · Cited by 49 · see citing works
Self-citationPochini, L., Galluccio, M., Scalise, M., Console, L., Pappacoda, G., & Indiveri, C. (2022) · International Journal of Molecular Sciences · doi:10.3390/ijms23020914 · Cited by 47 · see citing works
Nikodemus, D., Lazić, D., Bach, M. L., Bauer, T., Pfeiffer, C., Wiltzer, L., Lain, E., Schömig, E., & Gründemann, D. (2011) · PubMed · Cited by 40 · see citing works
Self-citationRegeneration of ergothioneine after reaction with singlet oxygen
Oumari, M., Goldfuss, B., Stoffels, C., Schmalz, H.-G., & Gründemann, D. (2019) · Free Radical Biology and Medicine · doi:10.1016/j.freeradbiomed.2019.01.043 · Cited by 39 · see citing works
Self-citationFaucher, Q., Alarcan, H., Marquet, P., & Guellec, C. B. (2020) · Journal of Clinical Medicine · doi:10.3390/jcm9082610 · Cited by 30 · see citing works
Ergothioneine and mitochondria: An important protective mechanism?
Fong, Z. W., Tang, R. M. Y., Cheah, I. K.-M., Leow, D. M.-K., Chen, L., & Halliwell, B. B. (2024) · Biochemical and Biophysical Research Communications · doi:10.1016/j.bbrc.2024.150269 · Cited by 29 · see citing works
The role of low molecular weight thiols in Mycobacterium tuberculosis
Emani, C. S., Gallant, J., Wiid, I. J. F., & Baker, B. (2019) · Tuberculosis · doi:10.1016/j.tube.2019.04.003 · Cited by 27 · see citing works
The Role of Ergothioneine in Red Blood Cell Biology: A Review and Perspective
Thomas, T. A., Francis, R. O., Zimring, J. C., Kao, J. P. Y., Nemkov, T. G., & Spitalnik, S. L. (2024) · Antioxidants · doi:10.3390/antiox13060717 · Cited by 27 · see citing works
Metabolome Changes during In Vivo Red Cell Aging Reveal Disruption of Key Metabolic Pathways
Jamshidi, N., Xu, X., von Löhneysen, K., Soldau, K., Mohney, R. P., Karoly, E. D., Scott, M., & Friedman, J. S. (2020) · iScience · doi:10.1016/j.isci.2020.101630 · Cited by 19 · see citing works
Nishiyama, M., Nakamichi, N., Yoshimura, T., Masuo, Y., Komori, T., Ishimoto, T., Matsuo, J., & Kato, Y. (2020) · Neurochemical Research · doi:10.1007/s11064-020-03118-8 · Cited by 17 · see citing works
The Natural Antioxidant Ergothioneine
Nguyen, T. H., Nagasaka, R., & Ohshima, T. (2013) · Elsevier eBooks · doi:10.1016/b978-0-9830791-6-3.50015-1 · Cited by 13 · see citing works
Wide tolerance to amino acids substitutions in the OCTN1 ergothioneine transporter
Frigeni, M., Iacobazzi, F., Yin, X., & Longo, N. (2016) · Biochimica et Biophysica Acta (BBA) - General Subjects · doi:10.1016/j.bbagen.2016.03.021 · Cited by 11 · see citing works
Organic cation transporter novel 1 (OCTN1): beyond an ergothioneine transporter
Guo, Y., Zhou, K., Zhang, J., & Tao, Z. (2026) · Cell Communication and Signaling · doi:10.1186/s12964-026-02907-y · Cited by 9 · see citing works
- Authors
- Bacher, P., Giersiefer, S., Bach, M., Fork, C., Schömig, E., & Gründemann, D.
- Affiliations
- University of Cologne (DE)
- Corresponding author
- Gründemann, Dirk
- ORCID
- Gründemann, Dirk
- Venue
- Biochimica et Biophysica Acta (BBA) - Biomembranes
- Publisher
- Elsevier BV
- Published
- 2009-12
- Volume
- 1788
- Issue
- 12
- Pages
- 2594-2602
- ISSN
- 0005-2736 (ISSN-L)
- OpenAlex
- W2033191772
- Cited by
- 27
- References
- 29
- Citations by year
- 2026: 1 · 2024: 3 · 2022: 1 · 2021: 2 · 2020: 4 · 2019: 3 · 2018: 1 · 2017: 1 · 2016: 2 · 2015: 1 · 2014: 2 · 2013: 1 · 2012: 1
- FWCI
- 0.83
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Licence
- Locations
- Biochimica et Biophysica Acta (BBA) - Biomembranes (publishedVersion)
- PubMed (publishedVersion)
- Funders
- Koeln Fortune Program, Faculty of Medicine, University of Cologne (157/2007 and 99/2008)
- Topics
- Health Sciences › Medicine › Oncology › Drug Transport and Resistance Mechanisms
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Biochemistry › Amino Acid Enzymes and Metabolism
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Biochemistry › Sulfur Compounds in Biology
- Keywords
- carnitine transporter, SLC22A4, SLC22A5, OCTN1, OCTN2, substrate discrimination, amino acid substitutions, gain-of-function mutations, carnitine transport, transporter conformational change, substrate binding site, transmembrane helices
- MeSH terms
- Solute Carrier Family 22 Member 5, Biological Transport, Carnitine, Cell Line, Ergothioneine, Humans, Protein Binding, Substrate Specificity, Sequence Homology, Amino Acid, Protein Structure, Tertiary, Antiporters, Amino Acid Substitution, Mutation, Missense, Organic Cation Transport Proteins
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
Are you an author, or do you see a problem with this page? Request a review
