Figure 8.
Effect of NGR-2C and isoDGR-2C on FN-RGE assembly. (A) Wild- type and FNRGE/RGE cells were cultured on LM111-coated dishes in serum replacement medium for 2 h, incubated with 800 μM NGR-2C or isoDGR-2C for 14 h, and stained with anti-FN antibodies. Note that both NGR-2C and isoDGR inhibit assembly of FN-RGE but not of wild-type FN. Bar, 15 μm. (B) Kinetics of NGR deamidation and rearrangement to isoDGR and DGR, respectively. NGR-2C peptide was incubated at 37°C and pH 7.2. Samples were taken the indicated times. Amounts of NGR and the rearrangement products isoDGR and DGR were determined with the HPLC as percentage peak areas. Results were normalized against NGR input. Note that the concentration of isoDGR is steadily increasing with time, reaching 10% of the NGR input after 16 h.