Characterization of high affinity binding motifs for the discoidin domain receptor DDR2 in collagen.
The discoidin domain receptors, DDR1 and DDR2, are receptor tyrosine kinases that are activated by native triple-helical collagen. Here we have located three specific DDR2 binding sites by screening the entire triple-helical domain of collagen II, using the Collagen II Toolkit, a set of overlapping triple-helical peptides. The peptide sequence that bound DDR2 with highest affinity interestingly contained the sequence for the high affinity binding site for von Willebrand factor in collagen III. Focusing on this sequence, we used a set of truncated and alanine-substituted peptides to characterize the sequence GVMGFO (O is hydroxyproline) as the minimal collagen sequence required for DDR2 binding. Based on a recent NMR analysis of the DDR2 collagen binding domain, we generated a model of the DDR2-collagen interaction that explains why a triple-helical conformation is required for binding. Triple-helical peptides comprising the DDR2 binding motif not only inhibited DDR2 binding to collagen II but also activated DDR2 transmembrane signaling. Thus, DDR2 activation may be effected by single triple-helices rather than fibrillar collagen.
Konitsiotis
Antonios D
AD
Division of National Heart and Lung Institute, Imperial College London, London SW7 2AZ.
Raynal
Nicolas
N
Bihan
Dominique
D
Hohenester
Erhard
E
Farndale
Richard W
RW
Leitinger
Birgit
B
D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D002417:Cattle; D002462:Cell Membrane / Q000378:metabolism; D003094:Collagen / Q000737:chemistry*; D000070818:Discoidin Domain Receptors; D006801:Humans; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D051381:Rats; D020794:Receptor Protein-Tyrosine Kinases / Q000737:chemistry*; D011975:Receptors, Mitogen / Q000737:chemistry*; D014841:von Willebrand Factor / Q000737:chemistry*
D010455:Peptides; D011975:Receptors, Mitogen; D014841:von Willebrand Factor; D003094:Collagen; D000070818:Discoidin Domain Receptors; D020794:Receptor Protein-Tyrosine Kinases