Table 2 Anisotropy data for secretin probes bound to WT and mutant SecR

Probes	Control		+GppNHp		Comparison Control vs +GppNHp, P values		Comparison WT vs mutant, P values
Probes	20 °C	37 °C	20 °C	37 °C	20 °C	37 °C	20 °C	37 °C
SecR WT
Alexa⁴⁸⁸-Sec	0.15 ± 0.01	0.08 ± 0.01	0.17 ± 0.01	0.07 ± 0.01	0.190	0.918
(Lys¹³-Alexa⁴⁸⁸)-Sec	0.14 ± 0.01	0.06 ± 0.01	0.20 ± 0.02*	0.12 ± 0.01*	0.046	0.044
(Lys²²-Alexa⁴⁸⁸)-Sec	0.14 ± 0.01	0.06 ± 0.01	0.20 ± 0.02**	0.12 ± 0.01*	0.004	0.011
Sec-Gly²⁸-(Cys²⁹-Alexa⁴⁸⁸)	0.13 ± 0.01	0.08 ± 0.01	0.13 ± 0.01	0.08 ± 0.01	0.999	0.999
SecR(G264A, I268A)
Alexa⁴⁸⁸-Sec	0.19 ± 0.01^#	0.1 ± 0.01	0.18 ± 0.01	0.1 ± 0.01	0.960	0.982	0.0163	0.266
(Lys¹³-Alexa⁴⁸⁸)-Sec	0.18 ± 0.02^#	0.11 ± 0.01	0.20 ± 0.01	0.11 ± 0.01	0.749	0.959	0.0477	0.102
(Lys²² -Alexa⁴⁸⁸)-Sec	0.18 ± 0.01^##	0.10 ± 0.01^#	0.19 ± 0.01	0.12 ± 0.01	0.635	0.229	0.005	0.0230
Sec-Gly²⁸-(Cys²⁹-Alexa⁴⁸⁸)	0.13 ± 0.01	0.09 ± 0.01	0.15 ± 0.01	0.1 ± 0.01	0.331	0.748	0.968	0.565

Values are expressed as means ± S.E.M. from three independent sets of observations analyzed using one-way ANOVA with Tukey post-test.
*P < 0.05 and **P < 0.01, significantly different from control at the same receptor construct, ^#P < 0.05 and ^##P < 0.01, significantly different from analogous condition at WT SecR.

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