X-ray structure of LeuT in an inward-facing occluded conformation reveals mechanism of substrate release

Kamil Gotfryd; Thomas Boesen; Jonas S. Mortensen; George Khelashvili; Matthias Quick; Daniel S. Terry; Julie W. Missel; Michael V. LeVine; Pontus Gourdon; Scott C. Blanchard; Jonathan A. Javitch; Harel Weinstein; Claus J. Loland; Poul Nissen; Ulrik Gether

doi:10.1038/s41467-020-14735-w

X-ray structure of LeuT in an inward-facing occluded conformation reveals mechanism of substrate release

Kamil Gotfryd, Thomas Boesen, Jonas S. Mortensen, George Khelashvili, Matthias Quick, Daniel S. Terry, Julie W. Missel, Michael V. LeVine, Pontus Gourdon, Scott C. Blanchard, Jonathan A. Javitch, Harel Weinstein, Claus J. Loland, Poul Nissen, Ulrik Gether

Publikation: Bidrag til tidsskrift/Konferencebidrag i tidsskrift /Bidrag til avis › Tidsskriftartikel › Forskning › peer review

31 Citationer (Scopus)

Abstract

Neurotransmitter:sodium symporters (NSS) are conserved from bacteria to man and serve as targets for drugs, including antidepressants and psychostimulants. Here we report the X-ray structure of the prokaryotic NSS member, LeuT, in a Na+/substrate-bound, inward-facing occluded conformation. To obtain this structure, we were guided by findings from single-molecule fluorescence spectroscopy and molecular dynamics simulations indicating that L-Phe binding and mutation of the conserved N-terminal Trp8 to Ala both promote an inward-facing state. Compared to the outward-facing occluded conformation, our structure reveals a major tilting of the cytoplasmic end of transmembrane segment (TM) 5, which, together with release of the N-terminus but without coupled movement of TM1, opens a wide cavity towards the second Na+ binding site. The structure of this key intermediate in the LeuT transport cycle, in the context of other NSS structures, leads to the proposal of an intracellular release mechanism of substrate and ions in NSS proteins.

Originalsprog	Engelsk
Artikelnummer	1005
Tidsskrift	Nature Communications
Vol/bind	11
Nummer	1
Antal sider	14
ISSN	2041-1723
DOI	https://doi.org/10.1038/s41467-020-14735-w
Status	Udgivet - 21 feb. 2020

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Gotfryd, K., Boesen, T., Mortensen, J. S., Khelashvili, G., Quick, M., Terry, D. S., Missel, J. W., LeVine, M. V., Gourdon, P., Blanchard, S. C., Javitch, J. A., Weinstein, H., Loland, C. J., Nissen, P., & Gether, U. (2020). X-ray structure of LeuT in an inward-facing occluded conformation reveals mechanism of substrate release. Nature Communications, 11(1), Artikel 1005. https://doi.org/10.1038/s41467-020-14735-w

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title = "X-ray structure of LeuT in an inward-facing occluded conformation reveals mechanism of substrate release",

abstract = "Neurotransmitter:sodium symporters (NSS) are conserved from bacteria to man and serve as targets for drugs, including antidepressants and psychostimulants. Here we report the X-ray structure of the prokaryotic NSS member, LeuT, in a Na+/substrate-bound, inward-facing occluded conformation. To obtain this structure, we were guided by findings from single-molecule fluorescence spectroscopy and molecular dynamics simulations indicating that L-Phe binding and mutation of the conserved N-terminal Trp8 to Ala both promote an inward-facing state. Compared to the outward-facing occluded conformation, our structure reveals a major tilting of the cytoplasmic end of transmembrane segment (TM) 5, which, together with release of the N-terminus but without coupled movement of TM1, opens a wide cavity towards the second Na+ binding site. The structure of this key intermediate in the LeuT transport cycle, in the context of other NSS structures, leads to the proposal of an intracellular release mechanism of substrate and ions in NSS proteins.",

author = "Kamil Gotfryd and Thomas Boesen and Mortensen, {Jonas S.} and George Khelashvili and Matthias Quick and Terry, {Daniel S.} and Missel, {Julie W.} and LeVine, {Michael V.} and Pontus Gourdon and Blanchard, {Scott C.} and Javitch, {Jonathan A.} and Harel Weinstein and Loland, {Claus J.} and Poul Nissen and Ulrik Gether",

year = "2020",

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day = "21",

doi = "10.1038/s41467-020-14735-w",

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Gotfryd, K, Boesen, T, Mortensen, JS, Khelashvili, G, Quick, M, Terry, DS, Missel, JW, LeVine, MV, Gourdon, P, Blanchard, SC, Javitch, JA, Weinstein, H, Loland, CJ, Nissen, P & Gether, U 2020, 'X-ray structure of LeuT in an inward-facing occluded conformation reveals mechanism of substrate release', Nature Communications, bind 11, nr. 1, 1005. https://doi.org/10.1038/s41467-020-14735-w

TY - JOUR

T1 - X-ray structure of LeuT in an inward-facing occluded conformation reveals mechanism of substrate release

AU - Gotfryd, Kamil

AU - Boesen, Thomas

AU - Mortensen, Jonas S.

AU - Khelashvili, George

AU - Quick, Matthias

AU - Terry, Daniel S.

AU - Missel, Julie W.

AU - LeVine, Michael V.

AU - Gourdon, Pontus

AU - Blanchard, Scott C.

AU - Javitch, Jonathan A.

AU - Weinstein, Harel

AU - Loland, Claus J.

AU - Nissen, Poul

AU - Gether, Ulrik

PY - 2020/2/21

Y1 - 2020/2/21

N2 - Neurotransmitter:sodium symporters (NSS) are conserved from bacteria to man and serve as targets for drugs, including antidepressants and psychostimulants. Here we report the X-ray structure of the prokaryotic NSS member, LeuT, in a Na+/substrate-bound, inward-facing occluded conformation. To obtain this structure, we were guided by findings from single-molecule fluorescence spectroscopy and molecular dynamics simulations indicating that L-Phe binding and mutation of the conserved N-terminal Trp8 to Ala both promote an inward-facing state. Compared to the outward-facing occluded conformation, our structure reveals a major tilting of the cytoplasmic end of transmembrane segment (TM) 5, which, together with release of the N-terminus but without coupled movement of TM1, opens a wide cavity towards the second Na+ binding site. The structure of this key intermediate in the LeuT transport cycle, in the context of other NSS structures, leads to the proposal of an intracellular release mechanism of substrate and ions in NSS proteins.

AB - Neurotransmitter:sodium symporters (NSS) are conserved from bacteria to man and serve as targets for drugs, including antidepressants and psychostimulants. Here we report the X-ray structure of the prokaryotic NSS member, LeuT, in a Na+/substrate-bound, inward-facing occluded conformation. To obtain this structure, we were guided by findings from single-molecule fluorescence spectroscopy and molecular dynamics simulations indicating that L-Phe binding and mutation of the conserved N-terminal Trp8 to Ala both promote an inward-facing state. Compared to the outward-facing occluded conformation, our structure reveals a major tilting of the cytoplasmic end of transmembrane segment (TM) 5, which, together with release of the N-terminus but without coupled movement of TM1, opens a wide cavity towards the second Na+ binding site. The structure of this key intermediate in the LeuT transport cycle, in the context of other NSS structures, leads to the proposal of an intracellular release mechanism of substrate and ions in NSS proteins.

UR - http://www.scopus.com/inward/record.url?scp=85079774084&partnerID=8YFLogxK

U2 - 10.1038/s41467-020-14735-w

DO - 10.1038/s41467-020-14735-w

M3 - Journal article

C2 - 32081981

AN - SCOPUS:85079774084

SN - 2041-1723

VL - 11

JO - Nature Communications

JF - Nature Communications

IS - 1

M1 - 1005

ER -

X-ray structure of LeuT in an inward-facing occluded conformation reveals mechanism of substrate release

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