Interaction proteomics of the human solute carrier (SLC) transporter superfamily

ABSTRACT: Solute carrier (SLC) transporters form a major superfamily which transport a wide range of substrates across cellular and organellar membranes. The superfamily consists of about 450 members and includes passive transporters, antiporters and symporters. To ensure the homeostatic regulation of metabolism and energetic household, transporters need to act in concert and be linked to the regulatory processes and pathways of cells. While there is ample evidence that regulation of transporter activity requires protein interactions, a comprehensive interactome of the whole human transproteome has not been described. We report here the first characterisation of the interactome of SLC transporters. To this aim, we applied an affinity purification combined with mass spectrometry (AP-MS) protocol tailored to transmembrane proteins to map the protein interactome for 400 solute carriers. The generated interactome covers thousands of novel protein-protein interactions, therefore providing a basis for understanding the molecular regulation of this protein superfamily. The SLC-interactome was dissected by clustering analysis, including SLC interactome-profile similarity-based clustering. The clustering revealed the association of SLCs with protein folding and trafficking. Thus, a selected set of interaction partners, associated with the proteostatic and trafficking regulation of SLCs, were selected for validation. We employed a high-throughput protein stability assay in combination with genetic perturbation of these selected interaction partners, to identify interactors which influence the abundance levels of SLCs or their subcellular locations. For selected interactions we performed transporter activity assays to show a direct modulation of transporter activity upon perturbation of protein interactions. Our work provides thus important insight into the complex molecular network of SLCs.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Kidney Cell

SUBMITTER: Fabian Frommelt  

LAB HEAD: Giulio Superti-Furga

PROVIDER: PXD055605 | Pride | 2025-03-03

REPOSITORIES: Pride