Figure 1
From: The transition state structure for coupled binding and folding of disordered protein domains
The experimental system.
(A) Example of an experimental binding trace for NCBDY2108W and an ACTR mutant, L1048A. Shown here is the main (fast) phase and protein concentrations are 1 μM NCBDY2108W and 10 μM L1048A-ACTR. (B) kobs values for the main phase obtained from binding traces (such as in panel A) plotted versus [ACTR] to obtain association rate constants, konapp by fitting the data to the general equation for the reversible association of two molecules under second order conditions46 (Eq. 4). At high concentrations of ACTR the equation approaches a linear function with a slope equal to kon. Red, NCBDY2108W and ACTRWT; blue, NCBDY2108W and ACTRL1048A. (C) Dissociation rate constants, koffapp, were determined in displacement experiments, in which NCBDWT was used to compete out NCBDY2108W from the complex. The solid line is a fit to a single exponential equation. At high concentration of NCBDWT the dissociation of NCBDY2108W/ACTR is virtually irreversible and kobs from the curve fitting is equal to koffapp. Red, NCBDY2108W/ACTRWT; blue, NCBDY2108W/ACTRL1048A. (D) Structure of the complex between ACTR (green) and NCBD (cyan), with L1048 highlighted in dark blue. The figure was generated using PyMol (The PyMol Molecular Graphics System, Version 1.3 Schrödinger, LLC).
