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J. Biol. Chem., Vol. 266, Issue 14, 8807-8813, May, 1991
F Fukai, H Suzuki, K Suzuki, A Tsugita and T Katayama
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.
Rat plasma fibronectin contains two distinct chemotactic domains for fibroblastic cells
Department of Patho-Physiology, Faculty of Pharmaceutical Sciences, Science University of Tokyo, Japan.
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