Clickable Substrate Transport (CST): A High-Throughput Functional Assay for Solute Carrier Proteins

Area of Science:

• Biochemistry and Molecular Biology
• Pharmacology
• Membrane Transport Proteins

Background:

• Solute carrier proteins (SLCs) are a large family of membrane proteins crucial for cellular function.
• Modulating SLCs holds therapeutic potential, but challenges exist in developing small molecules for less-studied members.
• Existing in vitro assays for SLCs are limited, hindering drug discovery efforts.

Purpose of the Study:

• To present a novel strategy for discovering and developing small molecules targeting various SLC transporters.
• To establish a robust and scalable cellular assay for monitoring substrate transport.
• To overcome limitations in current screening assays for challenging SLC targets.

Main Methods:

• Development of a cellular assay using a fluorogenic dye and bioorthogonal chemistry.
• Utilizing the glucose transporter SLC5a1 as a model system to validate the assay.
• Demonstrating the assay's robustness, scalability, and applicability across diverse substrates and targets.

Main Results:

• A novel cellular assay was successfully developed and validated using SLC5a1.
• The assay effectively reports on substrate transport through the application of a fluorogenic dye and bioorthogonal chemistry.
• The assay is robust, scalable, and adaptable for a wide range of SLC transporters.

Conclusions:

• The developed assay provides a powerful new tool for the discovery of small molecule modulators of SLC proteins.
• This work expands the limited catalog of screening assays for traditionally challenging transporter targets.
• The strategy is anticipated to accelerate the development of therapeutics targeting the therapeutically significant SLC superfamily.