EFFECT OF RECOMBINANT OSTEOPONTIN ON ADHESION AND MIGRATION OF P388D1 CELLS
EFFECT OF RECOMBINANT OSTEOPONTIN ON ADHESION AND MIGRATION OF P388D1 CELLS
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DOI：
10.1111/j.1749-6632.1995.tb44662.x
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发表时间：
1995-01-01
期刊：
影响因子：
--
通讯作者：
AKIZUKI, S
中科院分区：
文献类型：
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作者：
YAMAMOTO, S;NASU, K;AKIZUKI, S
Osteopontin (OPN) is the smallest natural Gly-Arg-Asp (RGD) protein yet identified, and its matrix adhesiveness may be much more limited than that of fibronectin (FN) which is approximately six times larger. The OPN structure around RGD amino acids is closely related to that of FN, and OPN has been reported to bind to a receptor integrin, a, &, that is also bound by FN, although FN also binds to other integrins. 1* 2 Therefore, it is possible that OPN modulates the function of FN. Here, we report the role of OPN in adhesion and haptotactic migration of a murine macrophage cell line, P388D1, using recombinant rabbit osteopontin (rerabOPN).Recombinant wild type rabbit osteopontin and the protein with an Asp-to-Glu transposition induced by a point mutation in the rabbit osteopontin (rabOPN) cDNA within the RGD sequence were expressed in E. coli and purified to homogeneity. P388D1 cells bind rerabOPN in a saturable manner. rerabOPN induced adhesion and haptotaxis of P388D1 cells, whereas mutated rabOPN (rOPmut) did not. Anti-rerabOPN IgG F (ab’), and synthetic GRGDS peptide inhibited rerabOPN-mediated adhesion and haptotaxis of P388D1 cells (TABLES 1 and 2). FN-mediated adhesion of P388D1 cells was markedly inhibited in the presence of fluid-phase rerabOPN. Adhesion of P388D1 cells to rerabOPN was significantly inhibited by anti-VLA, a (a,) and VLAS a (a5) monoclonal antibodies (mAbs), but not anti-vitronectin (VN) receptor a (a,) or Mac-la (a,) mAbs. Adhesion of P388D1 cells to FN and VN was significantly inhibited by anti-a, mAb but not anti-a,,-a5 or-aM mAbs. Haptotaxis of P388D1 cells to rerabOPN and VN was significantly inhibited by anti-a,, but not by anti-a,,-a5 and-ay rnAbs, whereas that to FN showed no inhibition with all three mAbs. Haptotaxis of P388D1 cells to VN was significantly inhibited with anti-a5 and anti-a, but not by anti-a, and-aM mAb. rerabOPN showed no chemotactic effect on P388D1 cells. Receptors other than a, & such as a&, and asp1 are likely to mediate the binding of rerabOP to P388D1 cells, although others may be involved, since inhibition with anti-a, and a5 mAbs was relatively weak. FN shows specific affinities for collagen, fibrin, heparin, DNA, glycosaminoglycans, and cell surfaces receptors. Although OPN has been suggested to bind positively charged matrix
