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. 2012 Jul 6;7(7):e39847. doi: 10.1371/journal.pone.0039847

Figure 2. Kinetic characteristics of SIRT6 deacetylase activity.

Figure 2

(A) Progress curves for purified, recombinant full-length wild-type (wt) and mutant SIRT6 proteins, expressed in E.coli. ▪ wt SIRT6 (2 µM), ▾H133W (10 µM), ▴ H133Y (10 µM), ○ N114A (10 µM), ◊ inhibition of wt (2 µM) with 20 mM NAM (this also generates an intrinsic level of background fluorescence). Error bars represent the standard error of mean duplicate values. (B) K m,app determination for peptide gave a value of 14.7+/−1.5 µM. Error bars represent the standard error of mean duplicate values. (C) K m,app determination for NAD+ gave a value of 52.4+/−18 µM. Error bars represent the standard error of mean duplicate values. (D) Deacetylation of purified H3/H4 histones with SIRT6. Deacetylation could be detected at a substrate:enzyme ratio of 1∶1.