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Cryo-EM studies of the human biogenic amine transporters


EMSL Project ID
50642

Abstract

The aims of the proposed research are to elucidate the first structure of the human dopamine transporter (hDAT) as well as to determine multiple structures of the human serotonin transporter (hSERT) in different conformational states, bound to substrate, inhibitors and allosteric modulators, by single particle cryo-EM. This work will have a major impact on the field of neurotransmitter transporters because, at present, it is difficult to elucidate the structures of these transporters by x-ray crystallography and the absence of structural information hinders the development of structure-based mechanisms and of understanding how small molecules, including widely prescribed antidepressants and mode stabilizing molecules, modulate transporter function. We have developed methods for large-scale expression, thermostabilizing mutants and 3D epitope-recognizing antibodies to enable structural studies of hDAT and hSERT. The use of transporter-Fab complexes is essential because they are small (ca. 60 kDa) membrane proteins and largely ensconced within the detergent micelle. We expect that our work will advance the transporter field by providing the first structure of hDAT as well as multiple new structures of hSERT in complexes with medically relevant and mechanistically important small molecules.

Project Details

Start Date
2019-01-14
End Date
2020-07-20
Status
Closed

Team

Principal Investigator

Eric Gouaux
Institution
Oregon Health & Science University

Team Members

Jonathan Coleman
Institution
University of Pittsburgh

Heidi Owen
Institution
Oregon Health & Science University

Dongxue Yang
Institution
Oregon Health & Science University

Vikas Navratna
Institution
Oregon Health & Science University

Janette Myers
Institution
Oregon Health & Science University

Irina El Khoury
Institution
Environmental Molecular Sciences Laboratory