Determinants at the N- and C-termini of Gα12 required for activation of Rho-mediated signaling

Table 1 Gα ₁₂ cassette mutants impaired in binding LARG-RH

70-100%	A-D F-H L N O T-V X BB DD-FF II KK XX YY BBB DDD III-KKK
40-70%	H P GG JJ WW ZZ CCC
20-40%	Y AA PP
10-20%	I OO QQ-SS UU VV FFF
0-10%	E J K M Q R S Z HH LL-NN TT AAA EEE GGG HHH
N/D	W CC

Cassette substitution mutants of myc-Gα₁₂ ^QL (see Figure 2 for alphabetical designations) were expressed in HEK293 cells and subjected to protein interaction assays using a GST-fusion of the RH domain of LARG as described in Methods, and for each mutant a pulldown:load ratio was determined and calculated as a percent (left column) of the same ratio for unmodified myc-Gα₁₂ ^QL assayed in parallel. Each Gα₁₂ mutant was analyzed in three independent experiments, except for mutants that appeared in the 70-100% category in two independent experiments.