The D2 period of collagen II contains a specific binding site for the human discoidin domain receptor, DDR2.
The human discoidin domain receptors (DDRs), DDR1 and DDR2, are expressed widely and, uniquely among receptor tyrosine kinases, activated by the extracellular matrix protein collagen. This activation is due to a direct interaction of collagen with the DDR discoidin domain. Here, we localised a specific DDR2 binding site on the triple-helical region of collagen II. Collagen II was found to be a much better ligand for DDR2 than for DDR1. As expected, DDR2 binding to collagen II was dependent on triple-helical collagen and was mediated by the DDR2 discoidin domain. Collagen II served as a potent stimulator of DDR2 autophosphorylation, the first step in transmembrane signalling. To map the DDR2 binding site(s) on collagen II, we used recombinant collagen II variants with specific deletions of one of the four repeating D periods. We found that the D2 period of collagen II was essential for DDR2 binding and receptor autophosphorylation, whereas the D3 and D4 periods were dispensable. The DDR2 binding site on collagen II was further defined by recombinant collagen II-like proteins consisting predominantly of tandem repeats of the D2 or D4 period. The D2 construct, but not the D4 construct, mediated DDR2 binding and receptor autophosphorylation, demonstrating that the D2 period of collagen II harbours a specific DDR2 recognition site. The discovery of a site-specific interaction of DDR2 with collagen II gives novel insight into the nature of the interaction of collagen II with matrix receptors.
Leitinger
Birgit
B
Department of Medicine, The Sackler Institute for Muscular Skeletal Research, University College London, 5 University Street, London WC1E 6JJ, UK. b.leitinger@ucl.ac.uk
Steplewski
Andrzej
A
Fertala
Andrzej
A
D001665:Binding Sites; D002460:Cell Line; D024043:Collagen Type II / Q000737:chemistry* / Q000378:metabolism*; D000070819:Discoidin Domain Receptor 1; D000070818:Discoidin Domain Receptors; D006801:Humans; D008024:Ligands; D010766:Phosphorylation; D011485:Protein Binding; D011487:Protein Conformation; D020794:Receptor Protein-Tyrosine Kinases / Q000235:genetics / Q000378:metabolism*; D011975:Receptors, Mitogen / Q000235:genetics / Q000378:metabolism*
D024043:Collagen Type II; D008024:Ligands; D011975:Receptors, Mitogen; C092030:DDR1 protein, human; D000070819:Discoidin Domain Receptor 1; D000070818:Discoidin Domain Receptors; D020794:Receptor Protein-Tyrosine Kinases