FIG. 8.

The binding of CNP to αvβ3 integrin is independent of the RGD-binding site. Left panel, Different amounts of RGD or HANTA peptides were added to a 96-well plate coated with αvβ3 integrin for 1 h at 37°C. After incubation, a fixed amount (1 mg/ml) of biotinylated CNP peptide was added, followed by incubation for 1 h at 37°C. The biotinylated peptide bound to the well was determined by a streptavidin-peroxidase assay. Right panel, An αvβ3 integrin coated plate was preincubated with the indicated amounts of CNP or RGD peptides, and a fixed amount (1 mg/ml) of biotinylated RGD peptide was then added, followed by incubation for 1 h at 37°C. The amount of biotinylated RGD peptide was determined by a streptavidin-peroxidase assay. The data are expressed as the percentage of the peptide bound when the wells were preincubated with PBS as a control. The arithmetic means and standard deviations of two independent experiments performed in duplicate are shown.