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In vivo crosslinking, cell fractionation, immunoprecipitation and MS identification of proteins crosslinked to E. coli GapA-3xFLAG

ABSTRACT: Our BATCH assay suggested that glycolytic enzymes could associate into complexes of varied stoichiometry and potentially interact with proteins outside of glycolysis, moreover, PfkA, FbaA and GapA were the top interacting proteins, showing strong interactions with both the PTS and PNPase. We constructed individual E. coli strains expressing chromosomally-encoded 3xFLAG-tagged variants of these enzymes (i.e., E. coli pfkA-3xFLAG, E. coli fbaA-3xFLAG and E. coli gapA-3xFLAG) and monitored the expression of the that the 3xFLAG-tagged proteins by western blotting. GapA was the most abundant of the proteins, followed by FbaA and PfkA, the E. coli gapA-3xFLAG strain was subsequently used for in vivo crosslinking and immunoprecipitation (CLIP-MS).

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ORGANISM(S): Escherichia Coli

SUBMITTER: Sue Lin-Chao

LAB HEAD: Sue Lin-Chao

PROVIDER: PXD068274 | Pride | 2026-07-16

REPOSITORIES: Pride

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