Chondroadherin, a leucine rich-repeat family member, contains a very C-terminal sequence CKFPTKRSKKAGRH359, now shown to bind to heparin with a KD of 13 µM. This observation led us to investigate if chondroadherin interacts via this C-terminal heparin-binding domain with glycosaminoglycan chains of proteoglycans at the cell surface. Cells was shown to bind this heparin binding peptide in FACS analysis and the interaction was to glycosaminoglycans as it was abolished when sulfation was inhibited by chlorate treatment of the cells. In separate experiments heparin and heparan sulfate inhibited the peptide interaction in a dose dependent manner. Using a human chondrosarcoma and a murine osteoblast cell line, ... More
Chondroadherin, a leucine rich-repeat family member, contains a very C-terminal sequence CKFPTKRSKKAGRH359, now shown to bind to heparin with a KD of 13 µM. This observation led us to investigate if chondroadherin interacts via this C-terminal heparin-binding domain with glycosaminoglycan chains of proteoglycans at the cell surface. Cells was shown to bind this heparin binding peptide in FACS analysis and the interaction was to glycosaminoglycans as it was abolished when sulfation was inhibited by chlorate treatment of the cells. In separate experiments heparin and heparan sulfate inhibited the peptide interaction in a dose dependent manner. Using a human chondrosarcoma and a murine osteoblast cell line, heparan sulfate proteoglycans were identified as the cell surface receptors involved in the binding. Different binding syndecans were identified in the two different cell lines, indicating that the same protein core of a proteoglycan may have structural and functional differences in the attached heparan sulfate chains. Upon binding to coated peptide cells spread, demonstrating engagement of the cytoskeleton, but no focal adhesion complex was formed. The number of cells adhering via their β1 integrin receptor to collagen type II or chondroadherin was profoundly and rapidly enhanced by addition of the heparin binding peptide. The peptide added to the cells caused ERK phosphorylation showing triggered intracellular signaling. The results show that heparan sulfate chain differ between various members of the proteoglycan families on a given cell, but also differ between the same proteoglycan on different cells with a potential for differential regulation of cellular activities.
