Revealing the location and dynamics of a concealed binding site in the dopamine transporter

Rong Zhu (), Walter Sandtner, Thomas Stockner, Alexander Heilinger, Marion Holy, Oliver Kudlacek, Linda Wildling, Kusumika Saha, Anna Sophie Fröhlich, Michael Bindl, Paraskevi Tziortzouda, Anna Haider, Julia Gobl, Saanfor Hubert Suh, Jawad Akbar Khan, Julia Bicher, Nina Kastner, Andreas Ebner, Hermann J. Gruber, Michael Freissmuth, Amy Hauck Newman (), Harald H. Sitte () and Peter Hinterdorfer ()
Additional contact information
Rong Zhu: Johannes Kepler University Linz
Walter Sandtner: Medical University of Vienna
Thomas Stockner: Medical University of Vienna
Alexander Heilinger: Johannes Kepler University Linz
Marion Holy: Medical University of Vienna
Oliver Kudlacek: Medical University of Vienna
Linda Wildling: Johannes Kepler University Linz
Kusumika Saha: Medical University of Vienna
Anna Sophie Fröhlich: Johannes Kepler University Linz
Michael Bindl: Johannes Kepler University Linz
Paraskevi Tziortzouda: Johannes Kepler University Linz
Anna Haider: Johannes Kepler University Linz
Julia Gobl: Johannes Kepler University Linz
Saanfor Hubert Suh: Johannes Kepler University Linz
Jawad Akbar Khan: Medical University of Vienna
Julia Bicher: Medical University of Vienna
Nina Kastner: Medical University of Vienna
Andreas Ebner: Johannes Kepler University Linz
Hermann J. Gruber: Johannes Kepler University Linz
Michael Freissmuth: Medical University of Vienna
Amy Hauck Newman: National Institute on Drug Abuse-Intramural Research Program
Harald H. Sitte: Medical University of Vienna
Peter Hinterdorfer: Johannes Kepler University Linz

Nature Communications, 2025, vol. 16, issue 1, 1-12

Abstract: Abstract The dopamine transporter (DAT) is linked to neuropsychiatric disorders including ADHD, Parkinson’s disease, and substance use disorders. Accordingly, DAT is the target of illicit drugs and clinically important medicines. However, the number and function of ligand binding sites in DAT is enigmatic due to conflicting data from available structures and molecular pharmacology. Herein, we design force sensors with DAT ligands and measure their interaction forces with wild-type and mutated DATs, from which two distinct populations of unbinding strengths and off-rates are detected. The high-force population is reduced by V152I and S422A mutations, or by substituting Na+ with K+ or NMDG+. In contrast, several modifications including mutation G386H, acetylation of K92 and K384, mutation K92A, mutation K384A, or protonation of H477 decrease the low-force population. The present data delineate the threshold of binding strength, which may account for certain ligand binding sites to be imperceptible in crystal or cryo-EM structures. Furthermore, the force spectra provide the information on the position and kinetic rates of a herein detected ligand binding site in DAT.

Date: 2025
References: Add references at CitEc
Citations:

Downloads: (external link)
https://www.nature.com/articles/s41467-025-59511-w Abstract (text/html)

Related works:
This item may be available elsewhere in EconPapers: Search for items with the same title.

Export reference: BibTeX RIS (EndNote, ProCite, RefMan) HTML/Text

Persistent link: https://EconPapers.repec.org/RePEc:nat:natcom:v:16:y:2025:i:1:d:10.1038_s41467-025-59511-w

Ordering information: This journal article can be ordered from
https://www.nature.com/ncomms/

DOI: 10.1038/s41467-025-59511-w

Access Statistics for this article

Nature Communications is currently edited by Nathalie Le Bot, Enda Bergin and Fiona Gillespie

More articles in Nature Communications from Nature
Bibliographic data for series maintained by Sonal Shukla () and Springer Nature Abstracting and Indexing ().