TABLE 1.

Kinetic parameters of LARG mutants binding to Gα_i/13 or Gα_i/13KA k_a, association rate constant; k_d, dissociation rate constant; K_D = k_d/k_a, equilibrium constant. Kinetic properties were estimated from BIAcore raw data (four to five serial dilutions for each protein) using BIAevaluation software, as indicated in Fig. 1C.

Protein	ALF	Gai/13			Gai/13KA
		ka	kd	KD	ka	kd	KD
		m–1 s–1	s–1	m	m–1 s–1	s–1	m
RDPC	–	NDa	ND	ND	ND	ND	ND
	+	4.4 × 105	3.0 × 10–4	6.8 × 10–10	2.9 × 105	6.5 × 10–3	2.6 × 10–8
RDP	–	ND	ND	ND	ND	ND	ND
	+	9.8 × 105	8.6 × 10–3	8.8 × 10–9	ND	ND	ND
DPC	–	ND	ND	ND	ND	ND	ND
	+	1.8 × 104	1.4 × 10–3	7.6 × 10–8	1.8 × 104	1.4 × 10–3	8.1 × 10–8
RH	–	ND	ND	ND	ND	ND	ND
	+	6.9 × 105	9.7 × 10–3	1.5 × 10–8	ND	ND	ND
DH/PH	–	2.7 × 104	2.8 × 10–2	1.1 × 10–6	3.7 × 104	3.2 × 10–2	8.5 × 10–7
	+	1.7 × 104	7.1 × 10–3	4.3 × 10–7	2.0 × 104	8.5 × 10–3	4.3 × 10–7
C	–	2.1 × 104	2.0 × 10–2	9.6 × 10–7	1.3 × 104	1.5 × 10–2	1.1 × 10–6
	+	2.4 × 104	8.6 × 10–3	3.6 × 10–7	9.4 × 103	9.9 × 10–3	1.0 × 10–6
PDZ	–	7.2 × 103	1.6 × 10–2	2.2 × 10–6	8.7 × 103	1.4 × 10–2	1.6 × 10–6
	+	5.0 × 103	1.3 × 10–2	2.5 × 10–6	3.9 × 103	1.1 × 10–2	2.9 × 10–6

^aND, the kinetic analysis was not done because the responses were less than 1/10 compared to those with Gα_i/13 in the presence of ALF^–