Intramolecular cross-linking in a bacterial homolog of mammalian SLC6 neurotransmitter transporters suggests an evolutionary conserved role of transmembrane segments 7 and 8

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Intramolecular cross-linking in a bacterial homolog of mammalian SLC6 neurotransmitter transporters suggests an evolutionary conserved role of transmembrane segments 7 and 8. / Kniazeff, Julie; Loland, Claus Juul; Goldberg, Naomi; Quick, Matthias; Das, Shonit; Sitte, Harald H; Javitch, Jonathan A; Gether, Ulrik.

In: Neuropharmacology, Vol. 49, No. 6, 11.2005, p. 715-23.

Research output: Contribution to journal › Journal article › Research › peer-review

Harvard

Kniazeff, J, Loland, CJ, Goldberg, N, Quick, M, Das, S, Sitte, HH, Javitch, JA & Gether, U 2005, 'Intramolecular cross-linking in a bacterial homolog of mammalian SLC6 neurotransmitter transporters suggests an evolutionary conserved role of transmembrane segments 7 and 8', Neuropharmacology, vol. 49, no. 6, pp. 715-23. https://doi.org/10.1016/j.neuropharm.2005.07.003

APA

Kniazeff, J., Loland, C. J., Goldberg, N., Quick, M., Das, S., Sitte, H. H., Javitch, J. A., & Gether, U. (2005). Intramolecular cross-linking in a bacterial homolog of mammalian SLC6 neurotransmitter transporters suggests an evolutionary conserved role of transmembrane segments 7 and 8. Neuropharmacology, 49(6), 715-23. https://doi.org/10.1016/j.neuropharm.2005.07.003

Vancouver

Kniazeff J, Loland CJ, Goldberg N, Quick M, Das S, Sitte HH et al. Intramolecular cross-linking in a bacterial homolog of mammalian SLC6 neurotransmitter transporters suggests an evolutionary conserved role of transmembrane segments 7 and 8. Neuropharmacology. 2005 Nov;49(6):715-23. https://doi.org/10.1016/j.neuropharm.2005.07.003

Author

Kniazeff, Julie ; Loland, Claus Juul ; Goldberg, Naomi ; Quick, Matthias ; Das, Shonit ; Sitte, Harald H ; Javitch, Jonathan A ; Gether, Ulrik. / Intramolecular cross-linking in a bacterial homolog of mammalian SLC6 neurotransmitter transporters suggests an evolutionary conserved role of transmembrane segments 7 and 8. In: Neuropharmacology. 2005 ; Vol. 49, No. 6. pp. 715-23.

Bibtex

@article{0aab568074c211dbbee902004c4f4f50,
title = "Intramolecular cross-linking in a bacterial homolog of mammalian SLC6 neurotransmitter transporters suggests an evolutionary conserved role of transmembrane segments 7 and 8",
abstract = "The extracellular concentration of the neurotransmitters dopamine, serotonin, norepinephrine, GABA and glycine is tightly controlled by plasma membrane transporters belonging to the SLC6 gene family. A very large number of putative transport proteins with a remarkable homology to the SLC6 transporters has recently been identified in prokaryotes. Here we have probed structural relationships in a 'microdoman' corresponding to the extracellular ends of transmembrane segments (TM) 7 and 8 in one of these homologs, the tryptophan transporter TnaT from Symbiobacterium thermophilum. We found that simultaneous - but not individual - substitution of Ala286 at the top of TM7 and Met311 at the top of TM8 with cysteines conferred sensitivity to submicromolar concentrations of Hg(2+) as assessed in a [(3)H]tryptophan uptake assay. Because Hg(2+) can cross-link pairs of cysteines, this suggests close proximity between TM 7 and 8 in the tertiary structure of TnaT as previously suggested for the mammalian counterparts. Furthermore, the inhibition of uptake upon cross-linking the two cysteines provides indirect support for a conserved conformational role of these transmembrane domains in the transport process. It was not possible, however, to transfer to TnaT binding sites for another metal ion, Zn(2+), that we previously engineered in the dopamine (DAT) and GABA (GAT-1) transporters between TM 7 and 8. This suggests that the structure of the TM7/8 microdomain is not identical with that of DAT and GAT-1. Hence, our data also emphasize possible structural differences that should be taken into account when interpreting future data on bacterial homologs of the SLC6 transporters.",
keywords = "Animals, Biological Evolution, Cross-Linking Reagents, Dose-Response Relationship, Drug, Escherichia coli, Extracellular Space, Gene Expression, Mammals, Membrane Transport Proteins, Mercury, Mesylates, Models, Molecular, Mutagenesis, Site-Directed, Protein Binding, Protein Structure, Quaternary, Sequence Homology, Amino Acid, Structure-Activity Relationship, Tritium, Tryptophan",
author = "Julie Kniazeff and Loland, {Claus Juul} and Naomi Goldberg and Matthias Quick and Shonit Das and Sitte, {Harald H} and Javitch, {Jonathan A} and Ulrik Gether",
year = "2005",
month = nov,
doi = "10.1016/j.neuropharm.2005.07.003",
language = "English",
volume = "49",
pages = "715--23",
journal = "Neuropharmacology",
issn = "0028-3908",
publisher = "Pergamon Press",
number = "6",

}

RIS

TY - JOUR

T1 - Intramolecular cross-linking in a bacterial homolog of mammalian SLC6 neurotransmitter transporters suggests an evolutionary conserved role of transmembrane segments 7 and 8

AU - Kniazeff, Julie

AU - Loland, Claus Juul

AU - Goldberg, Naomi

AU - Quick, Matthias

AU - Das, Shonit

AU - Sitte, Harald H

AU - Javitch, Jonathan A

AU - Gether, Ulrik

PY - 2005/11

Y1 - 2005/11

N2 - The extracellular concentration of the neurotransmitters dopamine, serotonin, norepinephrine, GABA and glycine is tightly controlled by plasma membrane transporters belonging to the SLC6 gene family. A very large number of putative transport proteins with a remarkable homology to the SLC6 transporters has recently been identified in prokaryotes. Here we have probed structural relationships in a 'microdoman' corresponding to the extracellular ends of transmembrane segments (TM) 7 and 8 in one of these homologs, the tryptophan transporter TnaT from Symbiobacterium thermophilum. We found that simultaneous - but not individual - substitution of Ala286 at the top of TM7 and Met311 at the top of TM8 with cysteines conferred sensitivity to submicromolar concentrations of Hg(2+) as assessed in a [(3)H]tryptophan uptake assay. Because Hg(2+) can cross-link pairs of cysteines, this suggests close proximity between TM 7 and 8 in the tertiary structure of TnaT as previously suggested for the mammalian counterparts. Furthermore, the inhibition of uptake upon cross-linking the two cysteines provides indirect support for a conserved conformational role of these transmembrane domains in the transport process. It was not possible, however, to transfer to TnaT binding sites for another metal ion, Zn(2+), that we previously engineered in the dopamine (DAT) and GABA (GAT-1) transporters between TM 7 and 8. This suggests that the structure of the TM7/8 microdomain is not identical with that of DAT and GAT-1. Hence, our data also emphasize possible structural differences that should be taken into account when interpreting future data on bacterial homologs of the SLC6 transporters.

AB - The extracellular concentration of the neurotransmitters dopamine, serotonin, norepinephrine, GABA and glycine is tightly controlled by plasma membrane transporters belonging to the SLC6 gene family. A very large number of putative transport proteins with a remarkable homology to the SLC6 transporters has recently been identified in prokaryotes. Here we have probed structural relationships in a 'microdoman' corresponding to the extracellular ends of transmembrane segments (TM) 7 and 8 in one of these homologs, the tryptophan transporter TnaT from Symbiobacterium thermophilum. We found that simultaneous - but not individual - substitution of Ala286 at the top of TM7 and Met311 at the top of TM8 with cysteines conferred sensitivity to submicromolar concentrations of Hg(2+) as assessed in a [(3)H]tryptophan uptake assay. Because Hg(2+) can cross-link pairs of cysteines, this suggests close proximity between TM 7 and 8 in the tertiary structure of TnaT as previously suggested for the mammalian counterparts. Furthermore, the inhibition of uptake upon cross-linking the two cysteines provides indirect support for a conserved conformational role of these transmembrane domains in the transport process. It was not possible, however, to transfer to TnaT binding sites for another metal ion, Zn(2+), that we previously engineered in the dopamine (DAT) and GABA (GAT-1) transporters between TM 7 and 8. This suggests that the structure of the TM7/8 microdomain is not identical with that of DAT and GAT-1. Hence, our data also emphasize possible structural differences that should be taken into account when interpreting future data on bacterial homologs of the SLC6 transporters.

KW - Animals

KW - Biological Evolution

KW - Cross-Linking Reagents

KW - Dose-Response Relationship, Drug

KW - Escherichia coli

KW - Extracellular Space

KW - Gene Expression

KW - Mammals

KW - Membrane Transport Proteins

KW - Mercury

KW - Mesylates

KW - Models, Molecular

KW - Mutagenesis, Site-Directed

KW - Protein Binding

KW - Protein Structure, Quaternary

KW - Sequence Homology, Amino Acid

KW - Structure-Activity Relationship

KW - Tritium

KW - Tryptophan

U2 - 10.1016/j.neuropharm.2005.07.003

DO - 10.1016/j.neuropharm.2005.07.003

M3 - Journal article

C2 - 16129457

VL - 49

SP - 715

EP - 723

JO - Neuropharmacology

JF - Neuropharmacology

SN - 0028-3908

IS - 6

ER -

ID: 68495