Figure 7:

Effect of VWD mutations on the hydrogen deuterium exchange (H → D) of the A1A2A3 domain (HEK293). A) 1 hour hydrogen deuterium exchange (H → D) of wild-type A1A2A3 (top and bottom panels, blue area), V1314D A1 (type 2B GOF VWD, top panel, red area), and F1369I A1 (type 2M LOF VWD, bottom panel, green area) as a function of residue position. HXMS was performed in triplicate at 1, 5, 10, 20 and 30 minutes, 1, 3 and 6 hours, and overnight time points (HXMS Supporting Information). Heavy black arrows indicate the location of the mutation in the sequence. Gray vertical areas represent the indicated secondary structure elements (top). Red vertical lines represent disulfide bonds. Magenta and green vertical lines indicate residues that are O-glycosylated and N-glycosylated respectively. Yellow vertical line represents the Y₁₆₀₅-M₁₆₀₆ ADAMTS13 scissile bond. B) Platelet adhesion to V1314D A1A2A3 (top) and lack of adhesion to F1369I (bottom). Platelets adhered to V1314D do not translocate (see supplemental movies). Shear rate is 1500 s⁻¹. C) HX fraction mapped onto the structures of the A1, A2, A3 domains (pdb IDs 1AUQ [61], 3GXB [64], and 1AO3 [65], respectively). Black = not resolved, blue = 0, white = 0.25, red ≥ 0.5. All structures are rendered using UCSF Chimera [63].