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Figure 1


Fig. 1. Vn binds PrPC in vitro and other PrPC ligands can compete for this interaction. (a) Autoradiogram of overlay assay. Indicated amounts of Ln, Fn, Vn, type IV collagen (Col) and BSA were adsorbed onto a membrane and allowed to bind to 125I-His6-PrPC. (b) PrPC-coated wells were incubated with 125I-Vn at the indicated concentrations. Wells were washed and radioactivity was measured. Scatchard plot is shown as an inset. The data represent mean ± s.d. (c) Competition assay in which 125I-Vn was incubated over PrPC-coated wells in the presence of increasing concentrations of unlabeled Vn, STI1, Ln or BSA. Bound 125I-Vn differed from addition of 5 µg unlabeled Vn (*P<0.001), addition of 10 µg unlabeled Vn or Ln (**P<0.001), or addition of 25 µg unlabeled Vn, Ln or STI1 (***P<0.001) according to Tukey's Test. (d) Competition assay in which 125I-Vn was incubated over PrPC-coated wells in the presence of increasing concentrations of unlabeled STI1 peptide or Ln {gamma}-1 peptide. The percentage of Vn binding was reduced in the presence of 20 or 40 µM STI1 peptide (*P<0.02, Student's t-test).





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