Je. Murphyullrich et al., SPARC MEDIATES FOCAL ADHESION DISASSEMBLY IN ENDOTHELIAL-CELLS THROUGH A FOLLISTATIN-LIKE REGION AND THE CA2-BINDING EF-HAND(), Journal of cellular biochemistry, 57(2), 1995, pp. 341-350
SPARC is a one of a group of extracellular matrix proteins that regula
te cell adhesion through a loss of focal adhesion plaques from spread
cells. We previously reported that SPARC reduced the number of bovine
aortic endothelial (BAE) cells positive for focal adhesions [Murphy-Ul
lrich et al. (1991): J Cell Biol 115:1127-1136]. We have now character
ized the effect of SPARC on the cytoskeleton of BAE cells. Addition of
SPARC to spread BAE cells caused a dose-dependent loss of focal adhes
ion-positive cells, that was maximal at similar to 1 mu g/ml (0.03 mu
M). Consistent with the loss of adhesion plaques as detected by interf
erence reflection microscopy, vinculin appeared diffuse and F-actin wa
s redistributed to the periphery of cells incubated with SPARC. Howeve
r, the distribution of the integrin alpha(v) beta(3) remained clustere
d in a plaque-like distribution. These data, and the observation that
SPARC binds to BAE cells but not to the extracellular matrix, indicate
that SPARC acts via interactions with cell surface molecules and not
by steric/physical disruption of integrin-extracellular matrix ligands
. To determine the region(s) of SPARC that mediate a loss of focal adh
esions, we tested peptides from the four distinct regions of SPARC. Th
e cationic, cysteine-rich peptide 2.1 (amino acids 54-73) and the Ca2-binding EF-hand-containing peptide 4.2 (amino acids 254-273) were act
ive in focal adhesion disassembly. Furthermore, antibodies specific fo
r these regions neutralized the focal adhesion-labilizing activity of
SPARC. These results are consistent with previous data showing that pe
ptide 2.1 and 4.2 interact with BAE cell surface proteins and indicate
that the loss of focal adhesions from endothelial cells exposed to SP
ARC is a receptor-mediated event. (C) 1995 Wiley-Liss, Inc.