Rat plasma fibronectin contains two distinct chemotactic domains for fibroblastic cells.
Mechanism of fibronectin (FN)-induced chemotaxis of fibroblastic cells has not been fully understood. The present study was performed to establish a molecular nature of the chemotactic region of rat plasma FN. The chemotactic dose-response pattern of intact FN for mouse embryo fibroblastic cells, NIH-L13 cells, which was represented as a "bell-shape" curve with a maximum activity at around 50 nM, changed to a "biphasic" mode through a proteolysis with thermolysin. Two distinct chemotactic components were isolated from the thermolytic fragments. One component, a fragment with a molecular mass of 110-150 kDa, was estimated to contain the central cell-binding domain and the carboxyl-terminal heparin-binding domain of the intact FN molecule. Cell migration stimulated by the 110-150-kDa fragment increased successively in a dose-dependent manner, and the capability to promote the migration was much higher than that of the intact FN (over 2-fold). The second chemotactic component, a fragment with a molecular mass of 21 kDa, was shown to reside in the carboxyl-terminal fibrin-binding domain. The 21-kDa fragment produced a bell-shape dose-response pattern, being consistent with the intact FN, whereas a maximum response occurred at a 100-fold lower concentration (0.5 nM) than that of the intact FN molecule. At higher concentrations, this fragment revealed an inhibitory activity for the cell migration in response to the 110-150-kDa fragment. No significant molecular interaction between these two active components was observed by polyacrylamide gel electrophoresis under nondenaturing conditions, suggesting that the 21-kDa fragment may act directly on the cell to inhibit the cell migration. These results suggest that rat plasma FN contains at least two chemotactically active components that regulate cooperatively chemotactic migration of fibroblastic cells.
Fukai
F
F
Department of Patho-Physiology, Faculty of Pharmaceutical Sciences, Science University of Tokyo, Japan.
Suzuki
H
H
Suzuki
K
K
Tsugita
A
A
Katayama
T
T
D000595:Amino Acid Sequence; D000818:Animals; D002630:Chemotactic Factors; D004305:Dose-Response Relationship, Drug; D005347:Fibroblasts / Q000166:cytology*; D005353:Fibronectins / Q000097:blood* / Q000737:chemistry; D066298:In Vitro Techniques; D051379:Mice; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010446:Peptide Fragments / Q000737:chemistry / Q000494:pharmacology; D051381:Rats; D013820:Thermolysin
D002630:Chemotactic Factors; D005353:Fibronectins; D010446:Peptide Fragments; D013820:Thermolysin