High-Throughput Expression and Purification of Human Solute Carriers for Structural and Biochemical Studies

Solute carriers, or SLCs, are responsible for uptaking nutrients and are involved in human diseases. However, there are few drugs targeting SLCs because they're poorly studied. We developed efficient approaches to clone, express, and purify SLCs.

Our workflow yields pure proteins, allowing biochemical and structural studies, and ultimately provides insights for drug discovery campaigns. With the numerous roles in biological processes, disease and drug delivery, SLCs have increasingly received the attention and become attractive therapeutical targets. The private, public ReSOLUTE IMI projects generated open access resources and data to enable novel discoveries in SLC function, physiology, pathology, and therapeutic targeting.

Traditionally, SLCs were challenging to study as they are integral membrane proteins with various substrates and diverse transport mechanisms. Recently, Cryo-EM, the BacMam expression system, and new solubilization reagents have led to a step change in their characterization. Pure protein samples are needed for biochemical and structural studies.

In human solute carriers often have a poor yield when over expressed and unstable when purified. Therefore, in the past, each target required years to develop individual structures to produce the protein sample for downstream applications. Our method is optimized for the parallel and cost-efficient study of multiple targets or constructs of a single target.

Therefore, it balances experimental flexibility, cost, time, and lab resources carefully. Plus, we offer open access to all our protocols and resources in this article and in the ReSOLUTE web portal.