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. Author manuscript; available in PMC: 2016 Oct 31.
Published in final edited form as: ACS Chem Biol. 2016 Mar 30;11(6):1552–1560. doi: 10.1021/acschembio.6b00083

Figure 5.

Figure 5

ADEP effects. a) BLI trajectories following transfer of ATP-stabilized ClpXP into ClpP-free buffer containing 2 mM ATP without or with 50 μM ADEP-2B. b) BLI trajectories following transfer of ATP-stabilized ClpXP into ClpP-free buffer containing 200 nM ADEP-2B and 2 mM ATP or ATPγS. c) ADEP-2B stimulation of dissociation. ATP-stabilized ClpXP was transferred into ClpP-free buffer containing 2 mM ATP and different ADEP-2B concentrations, and dissociation rate constants were determined by single-exponential fits. The line is a hyperbolic fit (R2 = 0.994) with values of 2.0 ± 0.13 s−1 for the maximum rate and 50 ± 7 μM for half-maximal stimulation. Sigmoidal equations for mechanisms involving two ADEPs (α2/(1+2α+α2); α = [ADEP]/Kμ; R2 = 0.971) or three ADEPs (α3/(1+3α+3α2+α3); R2 = 0.958) gave poorer fits. d) (top) ADEP-2B stimulation of decapeptide cleavage of 25 nM ClpP. The line is a hyperbolic fit (R2 = 0.995) with half-maximal inhibition at a total concentration of 240 ± 26 nM. (bottom) ADEP-2B inhibition of association of 200 nM ClpP. The line is a hyperbolic fit (R2 = 0.981) with half-maximal inhibition at a total concentration of 167 ± 20 nM.