Extended Data Fig. 1: Characterizing the binding mode of triazolopyridine-based compounds to katanin.
From: A chemical genetics approach to examine the functions of AAA proteins
(a-b) DSF traces (a) and changes in melting temperatures (∆Tm, b) for kata-AAA-WT in the presence of compounds 1-4 (500 µM, n = 2). As a reference, the corresponding trace for control is shown (5% DMSO, dashed line, data in Fig. 1d). (c) Four computational docking models for compound 1 (purple and blue, stick representation) bound to the katanin nucleotide-binding site (gray, ribbon representation). Variability hotspot residues are shown (stick representation, color-coded as in Fig. 1a). Other key amino acids in the katanin ATP-binding site are also shown (gray, stick representation). Pose 1 corresponds to the one in Fig. 2e. (d) Isothermal Titration Calorimetry (ITC)-based analyses of kata-AAA-WT in the presence of compounds 1-3. Raw injection heats are shown for titrations of compounds 1-3 against kata-AAA-WT (left panels), or compound titrations into buffer (right panels). Compound 4 could not be analyzed under similar conditions due to limited solubility. (e) Integrated data points and fitted binding curves used to determine Kd values. Data for graphs in (a-b) and (d-e) are available as source data.
