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Custom Vector Construction> | The nucleotide sequence of eIF2Bα (Uniprot accession: Q14232) was synthesized by Genscript with a C-terminal TEV-avi-FLAG tag and cloned into a pET45b vector (Novagen) for expression...Supernatants were purified by FLAG affinity chromatography (Genscript) | Get A Quote |
The multi-subunit translation initiation factor eIF2B is a control node for protein synthesis. eIF2B activity is canonically modulated through stress-responsive phosphorylation of its substrate eIF2. The eIF2B regulatory subcomplex is evolutionarily related to sugar-metabolizing enzymes, but the biological relevance of this relationship was unknown. To identify natural ligands that might regulate eIF2B, we conduct unbiased binding- and activity-based screens followed by structural studies. We find that sugar phosphates occupy the ancestral catalytic site in the eIF2Bα subunit, promote eIF2B holoenzyme formation and enhance enzymatic activity towards eIF2. A mutant in the eIF2Bα ligand pocket that causes Vanis... More