Blakely Lab

The Blakely Lab

Elucidation of Selective Permeation Pathways in Neurotransmitter Transporters

Members of the GAT/NET family of Na+ and Cl- coupled neurotransmitter transporter gene family (SLC6A) form selective permeation pathways for neurotransmitters, amino acids and nutrients across plasma membranes of neurons, glia and epithelial cells.

Although a twelve transmembrane topology is generally accepted (and supported by biochemical and X-ray crystal studies of a bacterial homolog), exactly where substrates pass through the membrane is still a topic of investigation. Understanding this fundamental aspect of transporter structure is key to elucidating how specificity for substrates is achieved and where antagonists act.

We have taken the approach of utilizing species-differences in substrate and antagonist recognition to identify sites contributing to the specificity of ligand recognition. These studies make use of transient and stable transfection of multiple species of transporter cDNAs (predominately encoding serotonin transporter (SERT) proteins), site-directed mutagenesis and site-specific modifying reagents in concert with functional studies that use radiotracer flux paradigms, radioligand binding methods, and biophysical analyses.

Finally, in collaboration with the lab of Jens Meiler, we are modeling the serotonin transporter (see movie below) to generate new hypotheses regarding structure and function that can be tested in living cells.