Screening of candidate substrates and coupling ions of transporters by thermostability shift assays.

ABSTRACT: Substrates of most transport proteins have not been identified, limiting our understanding of their role in physiology and disease. Traditional identification methods use transport assays with radioactive compounds, but they are technically challenging and many compounds are unavailable in radioactive form or are prohibitively expensive, precluding large-scale trials. Here, we present a high-throughput screening method that can identify candidate substrates from libraries of unlabeled compounds. The assay is based on the principle that transport proteins recognize substrates through specific interactions, which lead to enhanced stabilization of the transporter population in thermostability shift assays. Representatives of three different transporter (super)families were tested, which diffe

SUBMITTER: Majd H 

PROVIDER: S-EPMC6211832 | biostudies-literature | 2018 Oct

REPOSITORIES: biostudies-literature