Table 1. Isothermal titration calorimetry (ITC) measurements between B56γ and KIF4A.

B56γ12-380	Titrant	KD (µM)*	ΔH (kcal/mol)	TΔS (kcal/mol)
WT	KIF4A‡ WT	15.2 ± 0.1	−11.7 ± 0.7	−5.1 ± 0.7
WT	KIF4Abpm† (K1208A/K1209A/K1210A)	55.6 ± 16.8	−11.5 ± 2.2	−5.6 ± 2.4
WT	KIF4ALE (C1224L/S1225E)	0.32 ± 0.01	−10.0 ± 0.1	−2.0 ± 0.1
WT	KIF4ALE,PE (C1224L/S1225E/A1231P/H1232E)	0.10 ± 0.01	−13.1 ± 1.1	−3.6 ± 1.1
WT	KIF4ALE,PE,bpm (K1208A/K1209A/K1210A/C1224L/ S1225E/A1231P/H1232E)	0.22 ± 0.02	−10.7 ± 0.1	−1.6 ± 0.3
E310R D313R	KIF4ALE,PE (C1224L/S1225E/A1231P/H1232E)	0.19 ± 0.01	−11.7 ± 0.1	−2.5 ± 0.1
E276R E310R D313R	KIF4ALE,PE (C1224L/S1225E/A1231P/H1232E)	0.21 ± 0.01	−8.2 ± 0.4	−0.9 ± 0.3
WT	RM§ WT	0.13 ± 0.01	−6.0 ± 0.1	3.3 ± 0.1
WT	RMbpm (R563A/K564A/K565A/K566A)	0.28 ± 0.01	−7.5 ± 0.1	1.4 ± 0.1
WT	NHS¶WT	4.9 ± 0.9	−17.0 ± 2.2	−9.8 ± 2.1
WT	NHSbpm (R1618A/R1620A)	54.5 ± 16.9	−18.1 ± 2.7	−12.2 ± 2.5
WT	AIM1** WT	0.80 ± 0.09	−9.2 ± 0.4	−0.9 ± 0.5
WT	AIM1bpm (K716A/R717A/K718A/K719A/K721A)	14.9 ± 1.8	−8.2 ± 0.2	−1.6 ± 0.2

^*All reported measurements are performed with ITC buffer (50 mM sodium phosphate pH 7.5, 150 mM NaCl, 0.5 mM TCEP). Errors are from duplicate or triplicate measurements.

^† bpm, basic patch mutant.

^‡KIF4A variants, KIF4A_1192-1232.

^§RepoMan (RM) variants, RM_533-603.

^¶NHS variants, NHS_1616-1635.

^**AIM1 variants, AIM1_716-741.