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. 2011 Feb 1;286(14):12702–12711. doi: 10.1074/jbc.M110.166538

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© 2011 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 5. — Effects of α_T* (S43N) on the R*-dependent GDP/GTP exchange activity of wild-type α_T*. A, 50 μm GTPγS was added to 6 nm R*, 300 nm wild-type α_T*, 300 nm β1γ1, and varying concentrations of α_T*(S43N) bound to GDP. The tryptophan fluorescence emission of the sample was monitored in real time with the excitation wavelength set at 300 nm and the emission wavelength set at 345 nm. Rate constants obtained from single exponential fits of the data are plotted as a function of the concentration of added α_T*(S43N). Inset: intrinsic tryptophan fluorescence is plotted as a function of time at different concentrations of α_T*(S43N) (black: none, red: 6 nm, green: 46 nm). B, 50 μm GTPγS was added to 6 nm R*, 300 nm wild-type α_T*, 300 nm β1γ1, and varying concentrations of α_T*(S43N) bound to GTPγS. The tryptophan fluorescence emission of the sample was monitored in real time with the excitation wavelength set at 300 nm and the emission wavelength set at 345 nm. Rate constants obtained from single exponential fits of the data are plotted as a function of the concentration of added α_T*(S43N). Inset: intrinsic tryptophan fluorescence is plotted as a function of time at different concentrations of α_T*(S43N) (black: none, red: 47 nm, green: 188 nm). C, 50 μm GTPγS was added to a mixture containing either 5 nm (red or green line) or 115 nm R* (blue line), 300 nm wild-type α_T*, 300 nm β1γ1, in the presence (green or blue line) or absence (red line) of 30 nm α_T*(S43N) bound to GDP. The tryptophan fluorescence emission of the sample was monitored in real time with the excitation wavelength set at 300 nm and the emission wavelength set at 345 nm. D, 50 μm GTPγS was added to a mixture containing either 5 nm (red or green line) or 115 nm R* (blue line), 300 nm wild-type α_T*, 300 nm β1γ1, in the presence (green or blue line) or absence (red line) of 100 nm α_T*(S43N) bound to GTPγS. The tryptophan fluorescence emission of the sample was monitored in real time with the excitation wavelength set at 300 nm and the emission wavelength set at 345 nm.