Table 2 Biochemical analysis of the TrioC module.

DSF and GEF activation data for TrioC variants. ΔTm = Tm(variant) − Tm(WT). Fold GEF activation = average kobs(variant)/kobs(WT). Fold activation by Gαq·GDP·AlF4 = average kobs(variant + Gαq·GDP·AlF4)/kobs(variant + Gαq·GDP). Each variant was profiled in n = 3 experiments, each performed at least in technical duplicate. A one-way analysis of variance (ANOVA) with post Dunnett’s test was used to test for significance for ΔTm, fold GEF activation, and Gαq·GDP·AlF4 activation for each variant in comparison to WT. See Materials and Methods for further explanation of methods and statistical analysis used for this table.

VariantTm (°C)95% CI for ∆TmGEF activation
(fold/WT)
95% CI for GEF
activation
Fold activation by
q·GDP·AlF4
95% CI for fold
activation by
q·GDP·AlF4
WT0.0[−0.5,0.5]†1.0[0.9, 1.1]3.1[0.4, 5.9]
E2069A−6.0***[−7.3, −4.7]0.7[0.1, 1.3]NDND
M2146A−3***[−3.6, −2.4]0.8[−0.5, 2.2]NDND
S2208A−3.2***[−4.8, −1.7]1.4[0.1, 2.7]NDND
R2150A0.9*[−0.2, 1.9]0.9[0.03, 1.8]NDND
F2207A1.1[−0.6, 2.8]0.7[−0.1, 1.6]NDND
∆2204–2208−0.4[−1.1, 0.2]1.5[0.2, 2.8]NDND
∆2203–2209−0.5[−0.9, −0.1]1.0[−0.5, 2.7]NDND
G2149I−6.3***[−6.7, −5.9]1.9[−0.7, 4.6]NDND
E2069R/R2150E−7.9***[−14, −2.2]3.2**[1.6, 4.7]NDND
R2150E0.9[0.7, 1.1]1.0[−0.4, 2.4]NDND
2153ΔNDND3.0*[0.7, 5.3]NDND
2152ΔNDND11***[4.5, 18]NDND
2147ΔNDND14***[4.8, 22]NDND
2143ΔNDND0.2[−0.2, 0.6]NDND
G2149W−3.6***[−3.9, −3.3]4.3***[2.4, 6.4]1.5*[0.7, 2.3]
R2150Q−1.6***[−3.3, 0.2]4.5***[2.1, 6.9]2.2[1.2, 3.1]
R2150W−3.1***[−5.1, −1.2]9.3***[−0.4, 18]0.9**[0.7, 1.0]

†Data were collected using two experimental setups; the larger 95% confidence interval (CI) was chosen here.

*P < 0.05.

**P < 0.01.

***P < 0.005.