Skip to main content

Structural studies of a bacterial dicarboxylate transporter


EMSL Project ID
50679

Abstract

Human cells import citrate as an energy source through the sodium driven citrate transporter NaCT in the plasma membrane. A bacterial homolog of NaCT is the Vibro succinate transporter VcINDY. We aim to determine the structures of both proteins in various conformations using single particle cryo-EM. Both proteins have a molecular mass of ~55 kDa and form a dimer, with almost the entire molecule berried in the membrane. To increase the mass of the particle to be suitable for cryo-EM, we will generate conformational specific Fabs that bind to the proteins, yielding complexes of 150-200 kDa. Such complexes will be subjected to grid screening and data collection. For the bacterial VcINDY protein, we have already generated 17 types of Fabs, and grids for complexes with several Fabs are currently ready. For the human transporter NaCT we have already purified the transporter protein and are in the process of screening and purifying antibodies. In the first six months of the project, the main aim of the project will be grid screening followed by data collection

Project Details

Start Date
2019-03-15
End Date
2019-10-23
Status
Closed

Team

Principal Investigator

Da-Neng Wang
Institution
New York University School of Medicine

Team Members

Harry Scott
Institution
Oregon Health & Science University

David Sauer
Institution
New York University School of Medicine

Related Publications

Sauer et al. Structural basis for the reaction cycle of DASS dicarboxylate transporters eLife 2020 Sep 1;9:e61350. doi: 10.7554/eLife.61350.