653 / 2019-04-29 20:57:01

A global map of protein-metabolite interactions in A. thaliana and budding yeast S. cerevisiae

signaling,metabolites,protein-metabolite interactions

全体主题 > Omics& New Technology

Protein – metabolite interactions (PMIs) are pivotal for cell regulation and, therefore, enable life. Although their importance and in comparison to protein – protein, protein – metabolite complexes remain understudied. To fill this gap we have developed a method called PROMIS, which exploits size separation for the cell-wide, untargeted analysis of PMIs. Applying PROMIS to an A. thaliana and S. cerevisiae cell extract, we ascertained several known PMIs. We detected hundreds of metabolites separating with protein complexes, revealing an unprecedented complexity of the protein–metabolite interactome. By looking for similar elution behavior of metabolites and enzymes belonging to the same biochemical pathways, we identified putative feedback and feed-forward regulations in pantothenate biosynthesis, methionine salvage cycle and purine nucleoside salvage pathway. By interpolating dataset with the list of predicted PMIs retrieved from the STITCH database, we provided experimental validation for 225 PMIs of the predicted interactions in yeast. Next, by combining PROMIS with affinity purification and thermal proteome profiling, we delineated a list of high-confidence binders of Ser-Leu and Tyr-Asp, thereby implicating dipeptides in the control of amino acid metabolism, protein processing and glycolysis. We further demonstrated that Tyr-Asp and Ser-Leu affects the activity of the glycolytic enzymes A. thaliana GAPC and S. cerevisiae PGK1, respectively.