Collagen binding specificity of the discoidin domain receptors: binding sites on collagens II and III and molecular determinants for collagen IV recognition by DDR1.
The discoidin domain receptors, DDR1 and DDR2 are cell surface receptor tyrosine kinases that are activated by triple-helical collagen. While normal DDR signalling regulates fundamental cellular processes, aberrant DDR signalling is associated with several human diseases. We previously identified GVMGFO (O is hydroxyproline) as a major DDR2 binding site in collagens I-III, and located two additional DDR2 binding sites in collagen II. Here we extend these studies to the homologous DDR1 and the identification of DDR binding sites on collagen III. Using sets of overlapping triple-helical peptides, the Collagen II and Collagen III Toolkits, we located several DDR2 binding sites on both collagens. The interaction of DDR1 with Toolkit peptides was more restricted, with DDR1 mainly binding to peptides containing the GVMGFO motif. Triple-helical peptides containing the GVMGFO motif induced DDR1 transmembrane signalling, and DDR1 binding and receptor activation occurred with the same amino acid requirements as previously defined for DDR2. While both DDRs exhibit the same specificity for binding the GVMGFO motif, which is present only in fibrillar collagens, the two receptors display distinct preferences for certain non-fibrillar collagens, with the basement membrane collagen IV being exclusively recognised by DDR1. Based on our recent crystal structure of a DDR2-collagen complex, we designed mutations to identify the molecular determinants for DDR1 binding to collagen IV. By replacing five amino acids in DDR2 with the corresponding DDR1 residues we were able to create a DDR2 construct that could function as a collagen IV receptor.
National Heart and Lung Institute, Imperial College London, London, United Kingdom.
Huifang
H
Xu
Nicolas
N
Raynal
Stavros
S
Stathopoulos
Johanna
J
Myllyharju
Richard W
RW
Farndale
Birgit
B
Leitinger
D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D001665:Binding Sites; D024043:Collagen Type II / Q000378:metabolism*; D024061:Collagen Type III / Q000378:metabolism*; D024141:Collagen Type IV / Q000737:chemistry / Q000378:metabolism*; D000070818:Discoidin Domain Receptors; D057809:HEK293 Cells; D006801:Humans; D008969:Molecular Sequence Data; D010766:Phosphorylation; D011485:Protein Binding; D020794:Receptor Protein-Tyrosine Kinases / Q000819:agonists / Q000737:chemistry / Q000378:metabolism*; D011975:Receptors, Mitogen / Q000819:agonists / Q000737:chemistry / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000378:metabolism*; D016415:Sequence Alignment; D015398:Signal Transduction
D024043:Collagen Type II; D024061:Collagen Type III; D024141:Collagen Type IV; D011975:Receptors, Mitogen; D011993:Recombinant Fusion Proteins; D000070818:Discoidin Domain Receptors; D020794:Receptor Protein-Tyrosine Kinases