TY - JOUR
T1 - Thrombin binds to murine bone marrow-derived macrophages and enhances colony-stimulating factor-1-driven mitogenesis
AU - Clohisy, D. R.
AU - Erdmann, J. M.
AU - Wilner, G. D.
PY - 1990
Y1 - 1990
N2 - The binding and mitogenic properties of thrombin have been established in various transformed cell lines. In such systems, thrombin induces cell division in the absence of exogenous growth factors, and the enzyme is considered to act directly as a mitogen. This study explores thrombin's interaction with nontransformed, growth factor-dependent cells. Binding of 125I-α-thrombin to colony-stimulating factor (CSF)-1-dependent bone marrow-derived macrophages is saturable, time-dependent, and displaceable by both unlabeled α-thrombin, and esterolytically inactive thrombin. Both dissociation studies of pre-bound radio-labeled thrombin and Scatchard analysis assisted by the program 'Ligand' suggest adherence of thrombin-binding data to a multi-site model. There are an estimated 2 x 104 high affinity sites (K(d) = 7 x 10-9 M) and 2 x 106 low affinity sites (K(d) = 9 x 10-7 M) per cell. Quiescent bone marrow-derived macrophages were cultured with either 10-8 M thrombin, 1000 units of CSF-1/ml, or both and [3H]thymidine incorporation was determined. Thrombin alone did not induce mitogenesis. CSF-1 induced mitogenesis with peak [3H]thymidine incorporation occurring 24 h after addition of the mitogen. This CSF-1-dependent mitogenic influence was enhanced greater than 2-fold by treatment with thrombin.
AB - The binding and mitogenic properties of thrombin have been established in various transformed cell lines. In such systems, thrombin induces cell division in the absence of exogenous growth factors, and the enzyme is considered to act directly as a mitogen. This study explores thrombin's interaction with nontransformed, growth factor-dependent cells. Binding of 125I-α-thrombin to colony-stimulating factor (CSF)-1-dependent bone marrow-derived macrophages is saturable, time-dependent, and displaceable by both unlabeled α-thrombin, and esterolytically inactive thrombin. Both dissociation studies of pre-bound radio-labeled thrombin and Scatchard analysis assisted by the program 'Ligand' suggest adherence of thrombin-binding data to a multi-site model. There are an estimated 2 x 104 high affinity sites (K(d) = 7 x 10-9 M) and 2 x 106 low affinity sites (K(d) = 9 x 10-7 M) per cell. Quiescent bone marrow-derived macrophages were cultured with either 10-8 M thrombin, 1000 units of CSF-1/ml, or both and [3H]thymidine incorporation was determined. Thrombin alone did not induce mitogenesis. CSF-1 induced mitogenesis with peak [3H]thymidine incorporation occurring 24 h after addition of the mitogen. This CSF-1-dependent mitogenic influence was enhanced greater than 2-fold by treatment with thrombin.
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M3 - Article
C2 - 2186026
AN - SCOPUS:0025265561
VL - 265
SP - 7729
EP - 7732
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
SN - 0021-9258
IS - 14
ER -