Table 2.

The C-terminus of AGS3 inhibits GTPγS binding to Gα_i3

Time, min	GTPγS bound, % of total binding
	—	+AGS3424–650	+AGS3591–650
10	100	16.6	28.9
80	100	15.7	26.2

GTPγS was incubated in the absence or presence of 1 μM each of His-AGS3_424–650 or His-AGS3_591–650. Reactions were started by addition of 10 μl of Gα_i3-GDP (250 nM). Results represent the percentage of GTPγS binding remaining after 10 and 80 min of incubation. Binding of Gα_i3, defined as 100%, was 0.27 ± 0.01 after 10 min and 1.15 ± 0.04 pmol after 80 min. AGS3_424–650 (4 GoLocos) and AGS3_591–650 (last GoLoco) inhibit (84% and 74%, respectively) the binding of GTPγS to Gα_i3. n = 3.