. Author manuscript; available in PMC: 2013 Apr 24.

Published in final edited form as: Biochemistry. 2012 Apr 10;51(16):3451–3459. doi: 10.1021/bi300070z

TABLE 2.

Rates A1, A2, and A3-C1-C2 subunit generation during WT and P3-P3′ mutant FVIII activation by thrombin

Factor VIII	A1 subunit generation^a	A2 subunit generation^a	A3-C1-C2 subunit generation^a
	nM A1 min⁻¹ (nM IIa)⁻¹	nM A2 min⁻¹ (nM IIa)⁻¹	nM A3-C1-C2 min⁻¹ (nM IIa)⁻¹
WT	63 ± 0.9	21 ± 1.1	199 ± 29
372(P3-P3′)740	652 ± 42	236 ± 25	121 ± 7.1
372(P3-P3′)1689	175 ± 12	164 ± 22	123 ± 10
740(P3-P3′)372	147 ± 22	168 ± 16	124 ± 16
372(P3-P3′)740/R740Q	665 ± 19	157 ± 15	157 ± 13
372(P3-P3′)740/740(P3-P3′)372	576 ± 33	216 ± 22	99 ± 7.4

A1, A2, and A3-C1-C2 subunit generation rates by thrombin cleavage of WT and mutant FVIII were estimated by nonlinear least-squares regression analysis of the data shown in Figures 2–5. The data represent the average ± S.D. values of at least three separate experiments. Western blot assays were performed and data analyzed as described under “Experimental Procedures.”