Figure 3. Nrp^Chimera molecules exhibit reversed VEGF-A specificity.

(A) The secondary structure of WT Nrp and Nrp^Chimera was assessed by CD. The overlapping spectra of Nrp^Chimera with wild-type Nrp demonstrate that the incorporated mutations are not structurally deleterious. (B) Nrp1^Chimera (blue line) and Nrp2^Chimera (green line) were tested for their ability to selectively sequester AP-VEGF-A from Nrp1 adsorbed on affinity plates. The Nrp^Chimera molecules show reversed VEGF-A specificity with Nrp1^Chimera having a marked reduction in inhibitory potency (IC₅₀ = 62 µM) and Nrp2^Chimera exhibiting a significant gain in potency (IC₅₀ = 3.9 µM). Wild-type Nrp1 (black dotted line, IC₅₀ = 1.8 µM) and Nrp2 (grey dotted line, IC₅₀≈310 µM) are shown for comparison (data from Figure 1). Experiments were performed in triplicate and reported as the mean ±1 S.D.