{Reference Type}: Journal Article
{Title}: Discovery of metal-binding proteins by thermal proteome profiling.
{Author}: Zeng X;Wei T;Wang X;Liu Y;Tan Z;Zhang Y;Feng T;Cheng Y;Wang F;Ma B;Qin W;Gao C;Xiao J;Wang C;
{Journal}: Nat Chem Biol
{Volume}: 20
{Issue}: 6
{Year}: 2024 Jun 26
{Factor}: 16.174
{DOI}: 10.1038/s41589-024-01563-y
{Abstract}: Metal-binding proteins (MBPs) have various and important biological roles in all living species and many human diseases are intricately linked to dysfunctional MBPs. Here, we report a chemoproteomic method named 'metal extraction-triggered agitation logged by thermal proteome profiling' (METAL-TPP) to globally profile MBPs in proteomes. The method involves the extraction of metals from MBPs using chelators and monitoring the resulting protein stability changes through thermal proteome profiling. Applying METAL-TPP to the human proteome with a broad-spectrum chelator, EDTA, revealed a group of proteins with reduced thermal stability that contained both previously known MBPs and currently unannotated MBP candidates. Biochemical characterization of one potential target, glutamine-fructose-6-phosphate transaminase 2 (GFPT2), showed that zinc bound the protein, inhibited its enzymatic activity and modulated the hexosamine biosynthesis pathway. METAL-TPP profiling with another chelator, TPEN, uncovered additional MBPs in proteomes. Collectively, this study developed a robust tool for proteomic discovery of MBPs and provides a rich resource for functional studies of metals in cell biology.