CELL-SURFACE PHOSPHATIDYLINOSITOL-ANCHORED HEPARAN-SULFATE PROTEOGLYCAN INITIATES MOUSE MELANOMA CELL-ADHESION TO A FIBRONECTIN-DERIVED, HEPARIN-BINDING SYNTHETIC PEPTIDE

被引:79
作者
DRAKE, SL
KLEIN, DJ
MICKELSON, DJ
OEGEMA, TR
FURCHT, LT
MCCARTHY, JB
机构
[1] UNIV MINNESOTA,DEPT ORTHOPAED SURG BIOCHEM,MINNEAPOLIS,MN 55455
[2] UNIV MINNESOTA,DEPT PEDIAT,MINNEAPOLIS,MN 55455
[3] UNIV MINNESOTA,CTR BIOMED ENGN,MINNEAPOLIS,MN 55455
关键词
D O I
10.1083/jcb.117.6.1331
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
Cell surface heparan sulfate proteoglycan (HSPG) from metastatic mouse melanoma cells initiates cell adhesion to the synthetic peptide FN-C/H II, a heparin-binding peptide from the 33-kD A chain-derived fragment of fibronectin. Mouse melanoma cell adhesion to FN-C/H II was sensitive to soluble heparin and pretreatment of mouse melanoma cells with heparitinase. In contrast, cell adhesion to the fibronectin synthetic peptide CS1 is mediated through an alpha-4-beta-1 integrin and was resistant to heparin or heparitinase treatment. Mouse melanoma cell HSPG was metabolically labeled with [S-35]sulfate and extracted with detergent. After HPLC-DEAE purification, S-35-HSPG eluted from a dissociative CL-4B column with a K(av) approximately 0.45, while S-35-heparan sulfate (HS) chains eluted with a K(av) approximately 0.62. The HSPG contained a major 63-kD core protein after heparitinase digestion. Polyclonal antibodies generated against HSPG purified from mouse melanoma cells grown in vivo also identified a 63-kD core protein. This HSPG is an integral plasma membrane component by virtue of its binding to Octyl Sepharose affinity columns and that anti-HSPG antibody staining exhibited a cell surface localization. The HSPG is anchored to the cell surface through phosphatidylinositol (PI) linkages, as evidenced in part by the ability of PI-specific phospholipase C to eliminate binding of the detergent-extracted HSPG to Octyl Sepharose. Furthermore, the mouse melanoma HSPG core protein could be metabolically labeled with H-3-ethanolamine. The involvement of mouse melanoma cell surface HSPG in cell adhesion to fibronectin was also demonstrated by the ability of anti-HSPG antibodies and anti-HSPG IgG Fab monomers to inhibit mouse melanoma cell adhesion to FN-C/H II. S-35-HSPG and S-35-HS bind to FN-C/H II affinity columns and require 0.25 M NaCl for elution. However, heparitinase-treated I-125-labeled HSPG failed to bind FN-C/H II, suggesting that HS, and not HSPG core protein, binds FN-C/H II. These data support the hypothesis that a phosphatidylinositol-anchored HSPG on mouse melanoma cells (MPIHP-63) initiates recognition to FN-C/H II, and implicate PI-associated signal transduction pathways in mediating melanoma cell adhesion to this defined ligand.
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页码:1331 / 1341
页数:11
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