Extended Data Fig. 2: Engineering the active site of katanin to obtain an allele sensitized to covalent inhibitors.
From: A chemical genetics approach to examine the functions of AAA proteins
a, Primary sequence of the katanin N-loop motif, which contains D210. b, Partial sequence alignment of AAA proteins showing the residues in the N-loop motif (D210 is indicated by the arrow, alignment generated using Clustal Omega). c, Schematic showing the AAA domain (light gray box, not to scale), and the first and last residues of the ATPase active human katanin construct. The position of the D210C mutation is indicated by the red bar. d, Differential scanning fluorimetry of katanin-WT and katanin-D210C in the absence and presence of ADP (1 mM) (n = 2 independent experiments). One representative experiment is shown. e, SDS–PAGE gels of purified recombinant L211C and T250C X. laevis katanin mutant constructs (Coomassie blue staining). Data for the graph in (d) and the unmodified gel picture for (e) are available as source data.
