Direct assessment of substrate binding to the Neurotransmitter:Sodium Symporter LeuT by solid state NMR

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Direct assessment of substrate binding to the Neurotransmitter:Sodium Symporter LeuT by solid state NMR. / Erlendsson, Simon; Gotfryd, Kamil; Larsen, Flemming Hofmann; Mortensen, Jonas Sigurd; Geiger, Michel-Andreas; van Rossum, Barth-Jan; Oschkinat, Hartmut; Gether, Ulrik; Teilum, Kaare; Løland, Claus Juul.

I: eLife, Bind 6, e19314, 2017.

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningfagfællebedømt

Harvard

Erlendsson, S, Gotfryd, K, Larsen, FH, Mortensen, JS, Geiger, M-A, van Rossum, B-J, Oschkinat, H, Gether, U, Teilum, K & Løland, CJ 2017, 'Direct assessment of substrate binding to the Neurotransmitter:Sodium Symporter LeuT by solid state NMR' eLife, bind 6, e19314. https://doi.org/10.7554/eLife.19314

APA

Erlendsson, S., Gotfryd, K., Larsen, F. H., Mortensen, J. S., Geiger, M-A., van Rossum, B-J., ... Løland, C. J. (2017). Direct assessment of substrate binding to the Neurotransmitter:Sodium Symporter LeuT by solid state NMR. eLife, 6, [e19314]. https://doi.org/10.7554/eLife.19314

Vancouver

Erlendsson S, Gotfryd K, Larsen FH, Mortensen JS, Geiger M-A, van Rossum B-J o.a. Direct assessment of substrate binding to the Neurotransmitter:Sodium Symporter LeuT by solid state NMR. eLife. 2017;6. e19314. https://doi.org/10.7554/eLife.19314

Author

Erlendsson, Simon ; Gotfryd, Kamil ; Larsen, Flemming Hofmann ; Mortensen, Jonas Sigurd ; Geiger, Michel-Andreas ; van Rossum, Barth-Jan ; Oschkinat, Hartmut ; Gether, Ulrik ; Teilum, Kaare ; Løland, Claus Juul. / Direct assessment of substrate binding to the Neurotransmitter:Sodium Symporter LeuT by solid state NMR. I: eLife. 2017 ; Bind 6.

Bibtex

@article{1f2a1cf9bbc54799bbcd3947fb543a4d,
title = "Direct assessment of substrate binding to the Neurotransmitter:Sodium Symporter LeuT by solid state NMR",
abstract = "The Neurotransmitter:Sodium Symporters (NSSs) represent an important class of proteins mediating sodium-dependent uptake of neurotransmitters from the extracellular space. The substrate binding stoichiometry of the bacterial NSS protein, LeuT, and thus the principal transport mechanism, has been heavily debated. Here we used solid state NMR to specifically characterize the bound leucine ligand and probe the number of binding sites in LeuT. We were able to produce high-quality NMR spectra of substrate bound to microcrystalline LeuT samples and identify one set of sodium-dependent substrate-specific chemical shifts. Furthermore, our data show that the binding site mutants F253A and L400S, which probe the major S1 binding site and the proposed S2 binding site, respectively, retain sodium-dependent substrate binding in the S1 site similar to the wild-type protein. We conclude that under our experimental conditions there is only one detectable leucine molecule bound to LeuT.",
author = "Simon Erlendsson and Kamil Gotfryd and Larsen, {Flemming Hofmann} and Mortensen, {Jonas Sigurd} and Michel-Andreas Geiger and {van Rossum}, Barth-Jan and Hartmut Oschkinat and Ulrik Gether and Kaare Teilum and L{\o}land, {Claus Juul}",
year = "2017",
doi = "10.7554/eLife.19314",
language = "English",
volume = "6",
journal = "eLife",
issn = "2050-084X",
publisher = "eLife Sciences Publications Ltd.",

}

RIS

TY - JOUR

T1 - Direct assessment of substrate binding to the Neurotransmitter:Sodium Symporter LeuT by solid state NMR

AU - Erlendsson, Simon

AU - Gotfryd, Kamil

AU - Larsen, Flemming Hofmann

AU - Mortensen, Jonas Sigurd

AU - Geiger, Michel-Andreas

AU - van Rossum, Barth-Jan

AU - Oschkinat, Hartmut

AU - Gether, Ulrik

AU - Teilum, Kaare

AU - Løland, Claus Juul

PY - 2017

Y1 - 2017

N2 - The Neurotransmitter:Sodium Symporters (NSSs) represent an important class of proteins mediating sodium-dependent uptake of neurotransmitters from the extracellular space. The substrate binding stoichiometry of the bacterial NSS protein, LeuT, and thus the principal transport mechanism, has been heavily debated. Here we used solid state NMR to specifically characterize the bound leucine ligand and probe the number of binding sites in LeuT. We were able to produce high-quality NMR spectra of substrate bound to microcrystalline LeuT samples and identify one set of sodium-dependent substrate-specific chemical shifts. Furthermore, our data show that the binding site mutants F253A and L400S, which probe the major S1 binding site and the proposed S2 binding site, respectively, retain sodium-dependent substrate binding in the S1 site similar to the wild-type protein. We conclude that under our experimental conditions there is only one detectable leucine molecule bound to LeuT.

AB - The Neurotransmitter:Sodium Symporters (NSSs) represent an important class of proteins mediating sodium-dependent uptake of neurotransmitters from the extracellular space. The substrate binding stoichiometry of the bacterial NSS protein, LeuT, and thus the principal transport mechanism, has been heavily debated. Here we used solid state NMR to specifically characterize the bound leucine ligand and probe the number of binding sites in LeuT. We were able to produce high-quality NMR spectra of substrate bound to microcrystalline LeuT samples and identify one set of sodium-dependent substrate-specific chemical shifts. Furthermore, our data show that the binding site mutants F253A and L400S, which probe the major S1 binding site and the proposed S2 binding site, respectively, retain sodium-dependent substrate binding in the S1 site similar to the wild-type protein. We conclude that under our experimental conditions there is only one detectable leucine molecule bound to LeuT.

U2 - 10.7554/eLife.19314

DO - 10.7554/eLife.19314

M3 - Journal article

VL - 6

JO - eLife

JF - eLife

SN - 2050-084X

M1 - e19314

ER -

ID: 172467398