TABLE 3.

Apparent dissociation constants for EF-hand domain mutants binding to SUVs mimicking the phospholipid composition of the inner plasma membrane

The vesicle (SUV and LUV) composition was PE/PC/PS/PI/SM/CH equal to 34.5:10.5:21:4.5:4.5:25; the buffer was 20 mm Tris, 0.1 mm EGTA, pH 7.5.

EF constructs	Apparent KDa	KD (mutant)/KD (WT)
	mm
WT	0.31 ± 0.05	1.0
WT (LUVs)	0.33 ± 0.22
R150A/K151A/K154A	1.03 ± 0.19	3.3
R182A/K183A/R186A	0.81 ± 0.19	2.6
W144A	1.08 ± 0.36	3.5
L149A	0.45 ± 0.03	1.5
F162A	0.31 ± 0.01	1.0
F168A	1.54 ± 0.34	5.0
V176A	0.44 ± 0.02	1.4
Y180A	0.54 ± 0.06	1.7
F185A	0.46 ± 0.10	1.5
I201A	0.37 ± 0.03	1.2

^a This is the total concentration of phospholipids present for half of maximum binding. Errors were determined from the hyperbolic fit to the data.