Human keratinocytes adhere to a unique heparin-binding peptide sequence within the triple helical region of type IV collagen

The present studies were aimed at further characterizing the interaction between basement membrane molecules and normal cultured human keratinocytes because of the intimate association between basal keratinocytes and the basement membrane. The studies show that keratinocytes adhere to type IV collagen-coated substrata to a greater degree than substrata coated with similar concentrations of fibronectin and laminin. To further define cell-binding regions within type IV collagen, studies were performed using purified pepsin-generated triple helical fragments of type IV collagen and show that keratinocytes bind to sites within the triple-helical region of type IV collagen. To delineate specific cell adhesion promoting sequences, we studied a series of chemically synthesized peptides derived from the triple-helical region of type IV collagen. One peptide, designated Hep III, which is thirteen amino acids in length and binds heparin, was active in directly promoting keratinocyte adhesion. Furthermore, in competition assays, this peptide in solution was shown to inhibit keratinocyte adhesion to substrata coated with Hep III or intact type IV collagen. These studies show that keratinocytes bind directly to type IV collagen and chemically define a major cell-adhesion-promoting site within the triple-helical region.