Activation of platelet-transforming growth factor beta-1 in the absence of thrombospondin-1.
Thrombospondin-1 (TSP-1) has been shown to bind and activate transforming growth factor-beta1 (TGF-beta1). This observation raises the possibility that TSP-1 helps to sequester TGF-beta1 in platelet alpha granules and activates TGF-beta1 once both proteins are secreted. Herein, we evaluated the level of active and latent TGF-beta1 in the plasma and in the supernatant of thrombin-treated platelets from TSP-1 null and wild-type mice on two genetic backgrounds (C57BL/6 and 129Sv). The plasminogen activator inhibitor-1/luciferase bioassay and an immunological assay were used to determine active and latent TGF-beta1. No significant differences were observed in the levels of active and latent TGF-beta1 in the supernatant of thrombin-treated platelets from TSP-1 null and wild-type mice. Active and latent TGF-beta1 were significantly increased in the plasma and platelets of C57BL/6 mice as compared with 129Sv mice. In addition, there was an increase of plasma level of latent TGF-beta1 in TSP-1 null mice as compared with wild-type mice on the C57BL/6 background but not on the 129Sv background. No active TGF-beta1 was observed in the plasma of either TSP-1 null and wild-type mice. These data indicate that TSP-1 does not function as a chaperon for TGF-beta1 during platelet production and does not activate significant quantities of secreted TGF-beta1 despite a vast excess in the number of TSP-1 molecules as compared with TGF-beta1 molecules. Because platelet releasates from TSP-1 null mice contain active TGF-beta1, we suggest that other important mechanisms of physiological activation of TGF-beta1 probably exist in platelets.
Abdelouahed
M
M
Department of Pathology, Beth Israel Deaconess Medical Center and Harvard Medical School, Boston, Massachusetts 02215, USA.
Ludlow
A
A
Brunner
G
G
Lawler
J
J
D000818:Animals; D001792:Blood Platelets / Q000187:drug effects / Q000378:metabolism; D002478:Cells, Cultured; D004492:Edetic Acid / Q000494:pharmacology; D004797:Enzyme-Linked Immunosorbent Assay; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D011485:Protein Binding; D019700:Thrombospondin 1 / Q000502:physiology*; D016212:Transforming Growth Factor beta / Q000378:metabolism*
D019700:Thrombospondin 1; D016212:Transforming Growth Factor beta; D004492:Edetic Acid