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. 2015 Dec 3;291(7):3483–3495. doi: 10.1074/jbc.M115.680991

FIGURE 4.

FIGURE 4.

Analysis of ligand binding properties of GS by ITC. Glutamine, ATP, or AMP in concentrations as indicated were injected (45 × 6 μl) into 20 μm (A and B) or 10 μm (C–F) GS with stirring at 155 rpm. A, titration with 2 mm glutamine. Six (5.6 ± 0.16) binding sites were predicted from the titration data. B, titration with 2 mm ATP. Six (5.5 ± 0.13) binding sites were predicted. C, titration of GS in the presence of 2 mm ATP with 2 mm glutamine. The glutamine concentration at the maximum enthalpy (highest point of the curve) was 173 ± 34.7 μm. D, titration of GS in the presence of 0.5 mm glutamine with 5 mm ATP. The ATP concentration at the maximum enthalpy (lowest point of the curve) was 421 ± 31.2 μm. E, titration of GS in the presence of 2 mm ATP with 2 mm AMP. The AMP concentration at the maximum enthalpy (highest point of the curve) was 186 ± 14.2 μm. F, titration of GS in the presence of 0.5 mm AMP with 5 mm ATP. The ATP concentration at the maximum enthalpy (lowest point of the curve) was 23 ± 4.2 μm.