A Yeast‐Based Assay for Inhibitors of l ‐Lactate Transport Utilizing Fluorescent Biosensors
Year: 2024
Type: article
Abstract: Inhibitors of ʟ‐lactate transport are in development as a novel mode of action in antitumor therapy and malaria. Previously, we used radiolabeled ʟ‐lactate to assay transport via the human monocarboxylate transporter 1, MCT1, and the structurally unrelated malaria parasite's transporter, PfFNT. We encountered a sensitivity limit at IC50 around 100 nM possibly resulting from the required high cell number per sample. Here, we describe a sensitive background‐free high‐throughput assay in yeast based on fluorescent iLACCO biosensors. We used iLACCO for co‐expression and fusions with the transporter protein. Uptake of ʟ‐lactate produced strong intensiometric fluorescent responses that could be monitored in cell suspensions using a fluorometer and in individual cells by fluorescence microscopy. The signal decreased dose‐dependently in the presence of specific MCT1 and PfFNT inhibitors. Re‐evaluation of 36 PfFNT inhibitors yielded IC50 values below 100 nM now matching previous data on Ki compound affinity to isolated transporter protein.
Source: ChemMedChem
Authors: Finn Tiedjens, Maike Menzel, Pauline Stahnke, Hanna Grotewold, Cane Uzun, Derya Yildirim, Eric Beitz
Institution: Christian-Albrechts-Universität zu Kiel
Language: English
Subfield: Cancer Research
Domain: Life Sciences
Open Access status: hybrid
Estimated APC paid: $3,950
Funder: Deutsche Forschungsgemeinschaft
Awards: G1013375132, G2371348836
OpenAlex ID: W4405350672