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Isothermal calorimetric estimation of dissociation constants (Kd) for GPIbα binding to repeat 9 (A), repeat 12 (B), repeat 17 (C), repeat 19 (D), repeat 21 (E), and repeat 23 (F). All of the experiments were in degassed 25 mm sodium phosphate, pH 6.4, 5 mm NaCl, and 1 mm DTT at 30 °C. Note that repeat 4 was unstable, preventing the precise measurement of the affinity, but the quick HSQC experiment clearly shows that it binds to GPIbα (see Fig. 5A).
