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. Author manuscript; available in PMC: 2010 Aug 2.
Published in final edited form as: Biochemistry. 2006 Aug 1;45(30):9074–9084. doi: 10.1021/bi060567d

Figure 2.

Figure 2

Binding of mant-AMPPNP to PKR measured by forward anisotropy titration. The fluorescence anisotropy of 20 μM mant-AMPPNP was measured at increasing concentrations of PKR at an excitation wavelength of 360 nm and emission wavelength of 443 nm with spectral bandwiths of 2 nm and 4 nm for excitation and emission respectively. Fluorescence intensity at each polarizer orientation were integrated for 1 s and measurements were averaged until the maximum standard deviation drops below 0.1%. Each data point represents the average of 3–4 measurements. The solid line represents a fit of the data to a 1:1 binding model to obtain Kd = 33.8± 0.7μM, rL = 0.0213± 0.0003 and rEL = 0.1914± 0.0010.