Inhibition of collagen fibrillogenesis by cells expressing soluble extracellular domains of DDR1 and DDR2.
Collagen fiber assembly affects many physiological processes and is tightly controlled by collagen-binding proteins. However, to what extent membrane-bound versus cell-secreted collagen-binding proteins affect collagen fibrillogenesis is not well understood. In our previous studies, we had demonstrated that the membrane-anchored extracellular domain (ECD) of the collagen receptor discoidin domain receptor 2 (DDR2) inhibits fibrillogenesis of collagen endogenously secreted by the cells. These results led to a novel functional role of the DDR2 ECD. However, since soluble forms of DDR1 and DDR2 containing its ECD are known to naturally exist in the extracellular matrix, in this work we investigated if these soluble DDR ECDs may have a functional role in modulating collagen fibrillogenesis. For this purpose, we created mouse osteoblast cell lines stably secreting DDR1 or DDR2 ECD as soluble proteins. Transmission electron microscopy, fluorescence microscopy, and hydroxyproline assays were used to demonstrate that DDR ECD expression reduced the rate and quantity of collagen deposition and induced significant changes in fiber morphology and matrix mineralization. Collectively, our studies advance our understanding of DDR receptors as powerful regulators of collagen deposition in the ECM and elucidate their multifaceted role in ECM remodeling.
Flynn
Lisa A
LA
Davis Heart and Lung Research Institute, 473 West 12th Avenue, Columbus, OH 43210, USA.
Blissett
Angela R
AR
Calomeni
Edward P
EP
Agarwal
Gunjan
G
D016475:3T3 Cells; D000818:Animals; D001483:Base Sequence; D003094:Collagen / Q000737:chemistry* / Q000378:metabolism* / Q000648:ultrastructure; D017931:DNA Primers / Q000235:genetics; D000070819:Discoidin Domain Receptor 1; D000070818:Discoidin Domain Receptors; D005109:Extracellular Matrix / Q000737:chemistry / Q000648:ultrastructure; D006909:Hydroxyproline / Q000737:chemistry; D007700:Kinetics; D051379:Mice; D046529:Microscopy, Electron, Transmission; D008856:Microscopy, Fluorescence; D010006:Osteoblasts / Q000378:metabolism / Q000648:ultrastructure; D055503:Protein Multimerization; D017434:Protein Structure, Tertiary; D020794:Receptor Protein-Tyrosine Kinases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D011975:Receptors, Mitogen / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D014162:Transfection
D017931:DNA Primers; D011975:Receptors, Mitogen; D011994:Recombinant Proteins; D003094:Collagen; C083249:Ddr1 protein, mouse; D000070819:Discoidin Domain Receptor 1; D000070818:Discoidin Domain Receptors; D020794:Receptor Protein-Tyrosine Kinases; D006909:Hydroxyproline