Abstract
Fibronectin (FN) is a cell adhesion protein that binds integrins in a process also involving the protein-crosslinking enzyme transglutaminase 2 (TG2) as a co-receptor. The cell-adhesive property of TG2 has been linked to a complex formation with FN and to its ability to crosslink and polymerize FN on the cell surface. We tested here the effects of extracellular FN, before and after in vitro crosslinking and polymerization by TG2, on MC3T3-E1 osteoblast adhesion. We show that TG2-mediated crosslinking creates large, compacted chain-like protein clusters that include both TG2 and FN molecules as analyzed by Western blotting and atomic force microscopy. Crosslinking of FN significantly promotes osteoblast adhesion as measured by crystal violet staining, and enhances β1-integrin clustering on the cell surface as visualized by immunofluorescence microscopy. We hypothesize that TG2-mediated crosslinking enhances the cell-adhesive properties of FN by increasing the molecular rigidity of FN in the extracellular matrix.
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Acknowledgments
This study was supported by a grant to MTK from the Canadian Institutes of Health Research. MTK is a scholar of the Fonds de la recherche en santé du Québec, and a member of the McGill Center for Bone and Periodontal Research and the McGill Center for Biorecognition and Biosensors.
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Forsprecher, J., Wang, Z., Nelea, V. et al. Enhanced osteoblast adhesion on transglutaminase 2-crosslinked fibronectin. Amino Acids 36, 747–753 (2009). https://doi.org/10.1007/s00726-008-0125-7
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DOI: https://doi.org/10.1007/s00726-008-0125-7