EXPERIMENTAl.

CEI,L

RESEARCH

190, 17-24 (1990)

A Fibronectin-Related Synthetic Peptide, Pro-Ala-Ser-Ser, lnhibits Fibronectin Binding t6 the Cell Surface, Fibronectin-Promoted Cell Migration in Vitro, and Cell Migration in Viva HIDEKIKATOW,'SAIIAHARUYAZAWA,* Biology, Laboratory

and *Department

AND

SHOSUKE SOFUKU*

of Chemistry, Kikkyo llnivcwi~y,

Tokyo i 71. Japan

or Gly-Arg-Giy-Asp-Ser-Pro-Cys (GRGDSPC) [ 11, 141 in a manner similar to that seen in neural crest cell migration in the amphibian embryo (e.g., Ref. 141). In contrast, the binding of exogenous FN to the PMC surface is inhibited by such large synthetic decapeptides as Arg-Gly-Asp-Ser-Pro-Ala-Ser-Ser-Lys-Pro (RGDSPASSKP), but not by these smaller peptides [ 10, 111. Such previous observations suggest that the larger peptide may possess another secondary functional region, such as a cell-binding site, in addition to the RGDS amino acid sequence in the RGDSPASSKP peptide. For example, in chemotactic migration of 3T3 and SV3T3 cells, a larger peptide encompassing the RGDS amino acid sequence is chemotactic, but RGDS itself is not [l]. It also has been suggested that the secondary functional region may locate on the immediate carboxyl terminus side of the RGDS amino acid sequence, since no amino acid other than Gly is involved on the amino terminus side of the decapeptide [18]. In this study, four peptides [Pro-Ala (PA), Pro-AlaSer (PAS), Pro-Ala-Ser-Ser (PASS): and Pro-Ala-Ser-Ser-Lys (PASSK)] were synthesized and applied to PMCs in vitro and in uiuo. Each of these synthetic peptides comprises the immediate carboxyl terminus of RGDS in the RGDSPASSKP peptide [18] and has a different length in order to determine minimum functional size. On the basis of in uitro indirect immunohistochemistry, FN binding to the PMC surface was inhibited by PASS and PASSK, respectively, without, association of the RGDS amino acid sequence. FN-promoted PMC migration in vitro was also inhibited by P.4SS. PMC migration in uiuo was also inhibited by introduction of PASS into the blastocoel. On the other hand, PA and PAS did not inhibit any of the Ffi-related phenomena.

The biological activity of the amino acid sequence consisting of the immediate carboxyl terminus side of the Arg-Gly-Asp-Ser (RGDS) amino acid sequence in the cell-binding domain of intact fibronectin (FN) molecules was examined using synthetic peptides [RGDS, Gly-Arg-Gly-Asp-Ser-Pro (GRGDSP), Arg-GlyAsp-Ser-Pro-Ala-Ser-Ser-Lys-Pro (RGDSPASSKP), Pro-Ala (PA), Pro-Ala-Ser (PAS), Pro-Ala-Ser-Ser (PASS), and Pro-Ala-Ser-Ser-Lys (PASSK)]. These peptides were applied to the primary mesenchyme cells (PMCs) of the sea urchin, Clypeaster japonicus. In vitro immunohistochemistry indicated that the bindingof exogenous FN to the PMC surface was inhibited by the peptides RGDSPASSKP and PASS, but not by RGDS, GRGDSP, PA, or PAS. PASS and RGDS introduced into the blastocoel also inhibited PMC migration in vivo. FN-promoted PMC migration in vitro was also inbibited by PASS and RGDS. The present results indicate that the PASS peptide inhibits FN binding to the PMC surface and promotes PMC migration, suggesting that the FN molecule uses the PASS amino acid sequence to bind to the PMC surface and to promote PMC migration in the blastocoel. ,e IWO Academic PRSS. IDC. -

lNTR

A fibronectin-related synthetic peptide, Pro-Ala-Ser-Ser, inhibits fibronectin binding to the cell surface, fibronectin-promoted cell migration in vitro, and cell migration in vivo.

The biological activity of the amino acid sequence consisting of the immediate carboxyl terminus side of the Arg-Gly-Asp-Ser (RGDS) amino acid sequenc...
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