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. Author manuscript; available in PMC: 2013 Jan 24.
Published in final edited form as: Biochemistry. 2012 Jan 17;51(3):820–828. doi: 10.1021/bi201731p

Figure 5. Michaelis-Menten Analysis of FXase formed using FVIII LC mutants.

Figure 5

FVIII (1 nM) was activated by thrombin (30 nM) for 1 minute in the presence of 20 μM PSPCPE. FXa generation was initiated by the addition of FIXa (40 nM) and various concentrations of FX (0–500 nM) as described in Methods. The symbols used are as follows; wild type (open circles), Thr2012Ala (closed circles), Phe2014Ala (open squares), Thr2012Ala/Phe2014Ala (closed squares), and Thr2012Ala/Leu2013Ala/Phe2014Ala (open triangles). Experiments were performed at least three separate times and mean values are shown.