Recombinant Plg and the KKS. (A-B) Activation of FXII (A) or PK (B), 200 nM each, by 200 nM of Plm-Lys³¹¹ (blue), Plm-Glu³¹¹ (red), or vehicle (purple). Reactions without FXII or PK for Plg-Lys³¹¹ and Plg-Glu³¹¹ are indicated in green and gray, respectively. (C-E) BK generation in normal plasma (C), PK-deficient plasma (D), or FXII-deficient plasma (E) supplemented with 600 nM (final concentration) of Plg-Lys³¹¹ (blue) or Plg-Glu³¹¹ (red) after addition of tPA (125 nM final concentration). (F) BK generation in normal plasma after addition of ΔFXII (160 nM; lavender), ΔFXII and KV999272 (10 μM; steel blue), or vehicle (light blue). (G) BK generation in normal plasma supplemented with 600 nM of Plg-Glu³¹¹ (red and steel blue) or vehicle (lavender) in response to tPA (125 nM) in the absence (red) or presence (steel blue) of 10 μM of KV999272. (H) Activation of Plg-Lys³¹¹ (blue, light green, and orange) or Plm-Glu³¹¹ (red, purple, and gray), 200 nM each, by 20 nM of tPA (blue and red), PKa (light green and purple), or FXIIa (orange and gray). (I) BK generation in normal plasma supplemented with Plg-Lys³¹¹ (blue), Plg-Glu³¹¹ (red), or vehicle (purple) after addition of ΔFXII (final concentration, 62.5 nM). For reactions in panels A, B, and H, samples were removed at indicated times and protease measured by chromogenic assay. In panels C to G and I, samples were removed at indicated times and BK measured by ELISA. In panels A to H, error bars indicate standard errors of the mean for duplicate experiments, each with 2 separate measurements. In panel I, error bars indicate standard errors for 2 experiments.