The epithelial mitogen keratinocyte growth factor binds to collagens via the consensus sequence glycine-proline-hydroxyproline.
The binding of certain growth factors and cytokines to components of the extracellular matrix can regulate their local availability and modulate their biological activities. We show that mesenchymal cell-derived keratinocyte growth factor (KGF), a key stimulator of epithelial cell proliferation during wound healing, preferentially binds to collagens I, III, and VI. Binding is inhibited in a dose-dependent manner by denatured single collagen chains and collagen cyanogen bromide peptides. This interaction is saturable with dissociation constants of approximately 10(-8) to 10(-9) m and estimated molar ratios of up to three molecules of KGF bound to one molecule of triple helical collagen. Furthermore, collagen-bound KGF stimulated the proliferation of transformed keratinocyte or HaCaT cells. Ligand blotting of collagen-derived peptides points to a limited set of collagenous consensus sequences that bind KGF. By using synthetic collagen peptides, we defined the consensus sequence (Gly-Pro-Hyp)(n) as the collagen binding motif. We conclude that the preferential binding of KGF to the abundant collagens leads to a spatial pattern of bioavailable KGF that is dictated by the local organization of the collagenous extracellular matrix. The defined collagenous consensus peptide or its analogue may be useful in wound healing by increasing KGF bioactivity and thus modulating local epithelial remodeling and regeneration.
Ruehl
Martin
M
Department of Medicine I, Klinikum Benjamin Franklin, Free University of Berlin, Hindenburgdamm 30, 12200 Berlin, Germany.
Somasundaram
Rajan
R
Schoenfelder
Ines
I
Farndale
Richard W
RW
Knight
C Graham
CG
Schmid
Monika
M
Ackermann
Renate
R
Riecken
Ernst Otto
EO
Zeitz
Martin
M
Schuppan
Detlef
D
D020816:Amino Acid Motifs; D003094:Collagen / Q000737:chemistry / Q000378:metabolism*; D003101:Collodion / Q000737:chemistry; D003488:Cyanogen Bromide / Q000737:chemistry; D004305:Dose-Response Relationship, Drug; D051523:Fibroblast Growth Factor 7; D005346:Fibroblast Growth Factors / Q000378:metabolism*; D005998:Glycine / Q000737:chemistry; D006801:Humans; D006909:Hydroxyproline / Q000737:chemistry; D007700:Kinetics; D008024:Ligands; D010455:Peptides / Q000378:metabolism; D011392:Proline / Q000737:chemistry; D011485:Protein Binding; D011994:Recombinant Proteins / Q000378:metabolism; D015854:Up-Regulation
C496453:FGF7 protein, human; D008024:Ligands; D010455:Peptides; D011994:Recombinant Proteins; D051523:Fibroblast Growth Factor 7; D005346:Fibroblast Growth Factors; D003101:Collodion; D003094:Collagen; D011392:Proline; D003488:Cyanogen Bromide; D006909:Hydroxyproline; D005998:Glycine