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. Author manuscript; available in PMC: 2015 Mar 11.
Published in final edited form as: Nature. 2014 Jul 20;513(7517):251–255. doi: 10.1038/nature13557

Figure 4. FBP1 inhibits HIF independent of its enzymatic activity, through direct interaction with a HIFα “inhibitory domain”.

Figure 4

a, Crystal structure (PDB ID: 1EYJ) of porcine FBP1 in complex with AMP (blue) and fructose 6-phosphate (F6P, red). The N-terminal regulatory domain of FBP1 is coloured in green, and the C-terminal catalytic domain is coloured in violet. The G260 residue is highlighted in yellow. b, Growth of RCC10 cells ectopically expressing vector, FBP1, or FBP1 G260R in 1% serum medium. c, HIF reporter activity in RCC10 cells expressing vector, FBP1, FBP1 G260R, the regulatory domain of FBP1 (“R” domain), and catalytic domain of FBP1 (“C” domain). RCC10 cell lysates were immunoprecipitated with IgG, HIF1α antibody (d), or HIF2α antibody (e) and blotted for endogenous FBP1. f, GST pull-down analysis between recombinant FBP1 and recombinant GST or GST-tagged HIF1α. IB: immunoblot. g, GST pull-down analysis between recombinant HIF1α and recombinant GST-tagged, FBP1 exon truncations. h, GST pull-down analysis between recombinant FBP1 and GST-tagged HIF1α motifs. Values represent mean±s.d. (three experimental replicates). *p<0.01.