L

1 10 100 1000 Peptide concentration (nmol/ml)

A EXPERIMENTAL FIGURE 6-24 A specific tripeptide sequence (RGD) in the cell-binding region of fibronectin is required for adhesion of cells. The cell-binding region of fibronectin contains an integrin-binding heptapeptide sequence, GRDSPC in the single-letter amino acid code (see Figure 2-13). This heptapeptide and several variants were synthesized chemically. Different concentrations of each synthetic peptide were added to polystyrene dishes that had the protein immunoglobulin G (IgG) firmly attached to their surfaces; the peptides were then chemically cross-linked to the IgG. Subsequently, cultured normal rat kidney cells were added to the dishes and incubated for 30 minutes to allow adhesion. After the nonbound cells were washed away, the relative amounts of cells that had adhered firmly were determined by staining the bound cells with a dye and measuring the intensity of the staining with a spectrophotometer. The plots shown here indicate that cell adhesion increased above the background level with increasing peptide concentration for those peptides containing the RGD sequence but not for the variants lacking this sequence (modification underlined). [From M. D. Pierschbacher and E. Ruoslahti, 1984, Proc. Nat'l. Acad. Sci. USA 81:5985.]

Fibrin, heparan sulfate Collagen binding binding

Type I repeat Type II repeat Type III repeat

EIIIB EIIIA IIICS

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