H. Chen et al., PROPERTIES OF RECOMBINANT MOUSE THROMBOSPONDIN-2 EXPRESSED IN SPODOPTERA CELLS, The Journal of biological chemistry, 269(51), 1994, pp. 32226-32232
A baculovirus system was used to express full-length recombinant mouse
thrombospondin 2 (rTSP2) as a disulfide-bonded homotrimer with an NH2
terminus beginning with Asp(20). rTSP2, like TSP1, was more sensitive
to trypsin digestion if depleted of calcium ion. The trypsin digestio
n pattern of rTSP2 and TSP1 differed in that trypsin cut between the f
irst and second type 1 modules of rTSP2. For bovine aortic endothelial
cells adhering to TSP-coated polystyrene plates, reduction after coat
ing caused both TSPs to be much more adhesive; these adhesions were bl
ocked completely by RGDS peptide or antibody to alpha(v) beta(s) integ
rin. rTSP2 and TSP1 also mediated the adhesion of HT-29 human colon ad
enocarcinoma cells that carry alpha(v) beta(5) but not alpha(v) beta(3
) integrin. Antibody to alpha(v) beta(5) did not inhibit adhesion of H
T-29 cells to TSP1 or rTSP2. Rather, adhesion of HT-29 cells was decre
ased by treatment of TSPs with EDTA, abolished by reduction of the TSP
s, and, in the case of rTSP2, blocked by heparin. Adhesion of MG63 cel
ls to both TSPs was complex. Treatment with EDTA enhanced the adhesive
activity of rTSP2 but decreased the adhesive activity of TSP1. These
results show that TSP2 can be processed and secreted when overexpresse
d using baculovirus, TSP1 and rTSP2 differ in protease susceptibility
in the type 1 module region, and TSP1 and rTSP2 mediate cell adhesion
by complex and similar but not identical mechanisms.