Anti-CUB antibodies induce an open conformation and do not inhibit ADAMTS13 activity. (A) Activity of wtADAMTS13 in absence or presence of antibodies toward the FRETS-VWF73 substrate at pH 6 or pH 7.5 (n = 3). (B) Ratio of activity at pH 7.5 divided by activity at pH 6, as in indicator of conformation (n = 3). (C) Binding of wtADAMTS13 in the presence or absence of anti-CUB antibodies to the 6A6 antibody (n = 3). All binding is relative to wtADAMTS13 in the presence of the activating 17G2 antibody. (D) Activity of wtADAMTS13, in presence or absence of antispacer or anti-CUB antibodies, toward platelet-bound VWF multimers under flow (n = 2). Platelets bound to VWF multimers are counted over time and expressed as a percentage relative to the starting frame. Loss of platelets is the result of cleavage of the VWF multimer strings. FU,fluorescent unit; WT, wild-type ADAMTS13.
Figure 2.

Anti-CUB antibodies induce an open conformation and do not inhibit ADAMTS13 activity. (A) Activity of wtADAMTS13 in absence or presence of antibodies toward the FRETS-VWF73 substrate at pH 6 or pH 7.5 (n = 3). (B) Ratio of activity at pH 7.5 divided by activity at pH 6, as in indicator of conformation (n = 3). (C) Binding of wtADAMTS13 in the presence or absence of anti-CUB antibodies to the 6A6 antibody (n = 3). All binding is relative to wtADAMTS13 in the presence of the activating 17G2 antibody. (D) Activity of wtADAMTS13, in presence or absence of antispacer or anti-CUB antibodies, toward platelet-bound VWF multimers under flow (n = 2). Platelets bound to VWF multimers are counted over time and expressed as a percentage relative to the starting frame. Loss of platelets is the result of cleavage of the VWF multimer strings. FU,fluorescent unit; WT, wild-type ADAMTS13.

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