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. Author manuscript; available in PMC: 2018 Jun 28.
Published in final edited form as: Thromb Haemost. 2017 Apr 13;117(7):1358–1369. doi: 10.1160/TH17-01-0043

Figure 5.

Figure 5

Protein S-concentration dependence of FVa and FVIIIa degradation by APC derivatives. (A) FVa (2.5 nM) degradation by APC-WT (O) and APC-G74S (●) (1 nM each) in the presence of increasing concentrations of protein S (from 0 to 50 nM) was analyzed on PC/PS vesicles (25 μM) in TBS/Ca2+ for 1min. The remaining cofactor activity of FVa was determined by a prothrombinase assay (1nM FXa and 1μM prothrombin for 1 min). (B) Similar to A, except that FVIIIa (10 nM) degradation by APC-WT and APC-G74S (20 nM each) in the presence of increasing concentrations of protein S (from 0 to 500 nM) was analyzed on PC/PS vesicles (40 μM) in TBS/Ca2+ for 6 min. The remaining cofactor activity of FVIIIa was determined by an intrinsic Tenase (1 nM FIXa and 100 nM FX for 1 min) as described in Materials and methods. (C) The same as panel A, except that the PC/PS concentration-dependence of FVa (2.5 nM) degradation by APC-WT and APC-G74S (1 nM each) in the absence and presence of protein S (110 nM) was monitored.