The carboxy-terminal extension of the collagen binding domain of fibronectin mediates interaction with a 165 kDa membrane protein involved in odontoblast differentiation.
Terminal differentiation of the odontoblast is characterized by an elongation and a polarization of the cell. The change in the cell shape and the reorganization of the cytoplasm involve the microfilament system. An immunological approach has previously implicated a transmembrane interaction between fibronectin and vinculin in the control of odontoblast differentiation. A 165 kDa protein localized on the cell-surface of odontoblasts mediated this interaction. In order to define the nature of the interaction of the 165 kDa protein with fibronectin, peptides were prepared by proteolytic cleavage of fibronectin with alpha-chymotrypsin. The results indicate that the 165 kDa protein interacted with a 62 kDa peptide located towards the amino-terminal extremity of fibronectin, but not with a 47 kDa related fragment. Both these 62 kDa and 47 kDa peptides included the collagen-binding domain and were retarded on a heparin-Ultrogel column. Microsequences demonstrated that the 62 kDa and 47 kDa fragments had the same amino-terminal extremity and that the larger fragment was extended in the carboxy-terminal direction. This carboxy-terminal extension of the collagen binding domain of fibronectin is implicated in the interaction of this molecule with the 165 kDa protein. On the other hand, odontoblasts differentiated normally when tooth germs were cultured in the presence of GRGDS synthetic peptide, suggesting that RGD-dependent integrins were not involved in odontoblast differentiation. Staining of dental mesenchymal cells in primary culture and of differentiated odontoblasts in situ with antibodies directed against the beta 1-subunit of integrins confirmed previous observations and showed that although beta 1 integrins are involved in the attachment of cultured dental cells, they are not implicated in the process of odontoblast differentiation.
Lesot
H
H
Institut de Biologie Médicale, INSERM CJF 88-08, Faculté de Médecine, Strasbourg, France.
Fausser
J L
JL
Akiyama
S K
SK
Staub
A
A
Black
D
D
Kubler
M D
MD
Ruch
J V
JV
D008841:Actin Cytoskeleton / Q000502:physiology; D000595:Amino Acid Sequence; D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology; D015153:Blotting, Western; D002454:Cell Differentiation / Q000502:physiology; D002478:Cells, Cultured; D003094:Collagen / Q000378:metabolism* / Q000502:physiology; D004591:Electrophoresis, Polyacrylamide Gel; D005353:Fibronectins / Q000378:metabolism* / Q000502:physiology; D005455:Fluorescent Antibody Technique; D008565:Membrane Proteins / Q000378:metabolism / Q000502:physiology*; D008648:Mesoderm / Q000166:cytology; D051379:Mice; D008969:Molecular Sequence Data; D009804:Odontoblasts / Q000166:cytology* / Q000378:metabolism / Q000502:physiology; D009842:Oligopeptides / Q000502:physiology; D014083:Tooth Germ / Q000166:cytology / Q000502:physiology; D016596:Vinculin / Q000378:metabolism / Q000502:physiology
D000970:Antineoplastic Agents; D005353:Fibronectins; D008565:Membrane Proteins; D009842:Oligopeptides; D016596:Vinculin; D003094:Collagen; C049554:glycyl-arginyl-glycyl-aspartyl-serine