TABLE 1.

Summary of mantdADP binding to and release from acto-MV FlAsH in the presence (2 mM or 10 mM MgCl₂) and absence of Mg (4 mM EDTA).

Rate Constant	2 mM MgCl2				10 mM MgCl2
	4°C	10°C	15°C	25°C	4°C	10°C	15°C	25°C
k+ligand (μM−1•s−1)	1.56±0.13	2.65±0.05	3.16±0.09	5.55±0.23	1.21±0.07	2.00±0.11	2.49±0.15	4.72±0.21
ak−ligand (s−1) y-intercept	0.79±0.36	1.65±0.13	2.17±0.25	8.08±0.6	1.00±0.21	1.76±0.30	3.84±0.41	8.80±0.60
k−ligand (s−1)	0.70±0.20	1.70±0.01	4.50±0.10	12.60±0.10	0.68±0.01	1.59±0.01	3.04±0.01	7.86±0.03
1/Kligand (μM)	0.47±0.07	0.73±0.15	1.06±0.52	1.86±0.58	0.70±0.18	0.85±0.08	1.37±0.24	1.77±0.16
k+pocket (s−1)	0.24±0.15	0.19±0.02	0.74±0.39	1.33±0.91	0.11±0.01	0.37±0.29	0.40±0.08	0.74±0.43
k−pocket (s−1)	0.51±0.34	0.85±0.12	2.22±0.28	5.79±0.20	0.37±0.03	0.94±0.22	1.55±0.70	3.99±2.24
Kpocket = k+pocket/k−pocket	0.47±0.43	0.23±0.04	0.33±0.18	0.23±0.16	0.28 ±0.03	0.40 ±0.32	0.26±0.13	0.19±0.15
bKD (μM)	0.32±0.30	0.87±0.28	0.64±0.39	1.32±0.92	0.54±0.15	0.61±0.36	1.09±0.57	1.54±1.24
cKD (μM)	0.70±0.26	ND	0.92±0.16	0.98±0.32	ND	ND	ND	ND

Rate Constant	4 mM EDTA
	4°C	10°C	15°C	25°C
k+ligand (μM−1•s−1)	2.01±0.14	4.15±0.27	4.14±0.38	12.06±1.61
k−ligand (s−1) y-intercept	12.85±0.41	25.52±1.06	37.63±1.06	82.35±4.52
k−ligand (s−1)	14.1±0.1	25.3±0.2	42.2±0.6	108.9±0.8
1/Kligand (μM)	6.75±0.54	6.24±0.61	9.73±1.23	7.90±1.87
k+pocket (s−1)	ND	ND	ND	ND
k−pocket (s−1)	ND	ND	ND	d14.77 ±2.00
Kpocket = kon/koff	ND	ND	ND	ND
cKD (μM) (Fig. 3)	4.87±0.33	ND	3.87±1.01	2.55±1.17

^adetermined from the y-intercept of the linear fits in Figure 2

^bcalculated with equation 7

^cdetermined from the hyperbolic fits in Figure 3

^ddetermined from the fit of the data in Figure 6B (extrapolated to Mg-free conditions)