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. 2023 Feb 13;51(10):4701–4712. doi: 10.1093/nar/gkad045

Figure 2.

Figure 2.

Autoinhibition of HMGB1 accelerates the HMGB1-target association in the presence of decoys. (A) Autoinhibition of HMGB1 occurs via electrostatic interactions of its D/E repeats with two DNA binding domains and other positively charged regions (8–10). Due to the lack of the D/E repeats, the Δ30 variant does not undergo autoinhibition. The fluorescence-labeled probe DNA modified by a cisplatin is also shown. (B) Stopped-flow fluorescence experiments for kinetic investigations of protein-target association in the presence of decoys. (C) Time-course of FAM fluorescence anisotropy upon mixing of a protein solution with a DNA solution containing the probe DNA and tRNAs as decoys. The concentrations of the protein, probe DNA and tRNAs were 50, 10, and 8000 nM, respectively. (D) Protein concentration dependence of the apparent pseudo-first-order rate constant kapp for protein-target association in the presence of 8 μM tRNAs as decoys. The buffer was 10 mM potassium phosphate (pH 7.5), 1 mM DTT, 1 mM MgCl2 and 100 mM KCl.