A competitive inhibitor traps LeuT in an open-to-out conformation

Satinder K. Singh, Chayne L. Piscitelli, Atsuko Yamashita, Eric Gouaux

Research output: Contribution to journalArticle

300 Citations (Scopus)

Abstract

Secondary transporters are workhorses of cellular membranes, catalyzing the movement of small molecules and ions across the bilayer and coupling substrate passage to ion gradients. However, the conformational changes that accompany substrate transport, the mechanism by which a substrate moves through the transporter, and principles of competitive inhibition remain unclear. We used crystallographic and functional studies on the leucine transporter (LeuT), a model for neurotransmitter sodium symporters, to show that various amino acid substrates induce the same occluded conformational state and that a competitive inhibitor, tryptophan (Trp), traps LeuT in an open-to-out conformation. In the Trp complex, the extracellular gate residues arginine 30 and aspartic acid 404 define a second weak binding site for substrates or inhibitors as they permeate from the extracellular solution to the primary substrate site, which demonstrates how residues that participate in gating also mediate permeation.

Original languageEnglish (US)
Pages (from-to)1655-1661
Number of pages7
JournalScience
Volume322
Issue number5908
DOIs
StatePublished - Dec 12 2008

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tryptophan-leucine
Leucine
Plasma Membrane Neurotransmitter Transport Proteins
Ions
Aspartic Acid
Tryptophan
Arginine
Binding Sites
Amino Acids
Membranes

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Cite this

A competitive inhibitor traps LeuT in an open-to-out conformation. / Singh, Satinder K.; Piscitelli, Chayne L.; Yamashita, Atsuko; Gouaux, Eric.

In: Science, Vol. 322, No. 5908, 12.12.2008, p. 1655-1661.

Research output: Contribution to journalArticle

Singh, SK, Piscitelli, CL, Yamashita, A & Gouaux, E 2008, 'A competitive inhibitor traps LeuT in an open-to-out conformation', Science, vol. 322, no. 5908, pp. 1655-1661. https://doi.org/10.1126/science.1166777
Singh, Satinder K. ; Piscitelli, Chayne L. ; Yamashita, Atsuko ; Gouaux, Eric. / A competitive inhibitor traps LeuT in an open-to-out conformation. In: Science. 2008 ; Vol. 322, No. 5908. pp. 1655-1661.
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