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Internalization of basic fibroblast growth factor (bFGF) in cultured endothelial cells: Role of the low affinity heparin‐like bFGF receptors
Author(s) -
Rusnati Marco,
Urbinati Chiara,
Presta Marco
Publication year - 1993
Publication title -
journal of cellular physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.529
H-Index - 174
eISSN - 1097-4652
pISSN - 0021-9541
DOI - 10.1002/jcp.1041540119
Subject(s) - internalization , basic fibroblast growth factor , intracellular , receptor , microbiology and biotechnology , cell culture , growth factor , biology , chemistry , biochemistry , genetics
We have shown (Presta et al., Cell Regul., 2:719–726, 1991) that a long‐lasting interaction of basic fibroblast growth factor (bFGF) with endothelial GM 7373 cells is required to induce cell proliferation. In the present work, we have investigated the interaction of 125 I‐bFGF with GM 7373 cells, its pathway of internalization, and its intracellular fate under the same experimental conditions previously utilized to assess the mitogenic activity of the growth factor. Cell cultures were incubated with 10 ng/ml 125 I‐bFGF for 2 h at 4°C. Then, cells were shifted to 37°C without changing the medium. A rapid down‐regulation of high affinity sites, paralleled by a rapid internalization of 125 I‐bFGF, was observed during the first 1–2 h at 37°C. After 6–8 h, also low affinity sites down‐regulate. This was paralleled by a continuous internalization of 125 I‐bFGF and by a slow disappearance of the growth factor from the culture medium. This suggests that GM 7373 cells activate, when exposed to bFGF for a long period of time, a late internalization pathway mediated by low affinity sites. This hypothesis was supported by the following experimental evidence: 1) soluble heparin inhibited the prolonged internalization of 125 I‐bFGF and its binding to low affinity sites with the same potency; 2) treatment of GM 7373 cells with heparinase, which removes most of the low affinity sites, also inhibited the prolonged internalization of 125 I‐bFGF internalized via low affinity sites was partially protected from lysosomal degradation. This was the case also when 125 I‐bFGF was internalized in the presence of soluble heparin, suggesting that the complexes bFGF—surface glycosaminoglycan and bFGF—soluble heparin are maintained during the internalization of the growth factor. Moreover, the capacity of soluble heparin to inhibit the mitogenic activity of bFGF also when added to cell cultures several hours after the growth factor indicates that the requirement for a prolonged interaction of bFGF with GM 7373 cells in order to induce cell proliferation might be related to the late internalization of the growth factor via low affinity sites. © 1993 Wiley‐Liss, Inc.