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. 1999 Jan 1;13(1):76–86. doi: 10.1101/gad.13.1.76

Figure 6.

Figure 6

Interaction between the Neh2 domain of Nrf2 and the DGR domain of Keap1. (A) Schematic representations of the wild-type and mutant Gal4 DBD–Neh2 fusion proteins (B1 to B4) and the Gal4 AD–Keap1 fusion proteins (L1–L3). Construct B1 contains full length Neh2. B4 is the same construct as the Neh2 deletion mutant of Nrf2 in Fig. 3. B2 contains two amino acid substitutions in Neh2, whereas B3 lacks the amino-terminal 15 residues. L2 and L3 lack the DGR or BTB domain of Keap1, respectively. (B) Expression plasmids for various Gal4 DBD and Gal4 AD fusion proteins were introduced into the reporter yeast strain in the given combinations. The resulting transformants were tested for the His+ phenotype by spotting onto His− or His+ plates. (C) Results of the BIAcore interaction assay. Ligand (GST–Neh2, amino acids 1–73, shaded bars) or GST alone (open bars), and analyte (MBP–Keap1–DGR, amino acids 308–624) binding was tested at three different concentrations. The test and the control values, each carried out in triplicate, are shown with the s.e.