Identification of cell surface binding sites for PC-cell-derived growth factor, PCDGF, (epithelin/granulin precursor) on epithelial cells and fibroblasts

Biochem Biophys Res Commun. 1998 Apr 17;245(2):539-43. doi: 10.1006/bbrc.1998.8498.

Abstract

PC cell derived growth factor (PCDGF) is an 88-kDa glycoprotein purified from the culture medium of the highly tumorigenic mouse teratoma-derived cell line PC. PCDGF was shown to stimulate the proliferation of 3T3 fibroblasts and PC cells. Amino acid sequencing of PCDGF indicated its identity to the precursor for the 6-kDa polypeptides epithelins and granulins. In this paper, we investigated the binding of PCDGF to the mink lung epithelial cell line CCL64. Scatchard analysis indicates that 125I-PCDGF binding to CCL64 cells is curvilinear, corresponding to the existence of two classes of binding sites: high affinity binding sites (560 +/- 170 sites/cell) with a Kd1 of 43 +/- 15 pM and low affinity binding sites (16,350 +/- 5900 sites/cell) with a Kd2 of 3.9 +/- 1.9 nM. 125I-PCDGF was chemically crosslinked to cell surface receptors on CCL64 cells with disuccinimidyl suberate. A major crosslinked band of about 190 kDa with radiolabeled PCDGF was detected after SDS-PAGE, suggesting the presence of PCDGF binding sites with molecular weight of about 120 kDa. 125I. PCDGF crosslinking studies indicate the presence of PCDGF binding sites with a molecular weight similar to those of binding sites on CCL64 cells on the surface of two other PCDGF-responsive cell lines, 3T3 fibroblasts and PC cells. These data suggest that the receptors for PCDGF are widely distributed on cells of distinct embryonic origin.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Affinity Labels / chemistry
  • Animals
  • Binding Sites
  • Cell Line
  • Cross-Linking Reagents / metabolism
  • Gene Expression Regulation / genetics
  • Glycoproteins / metabolism*
  • Growth Substances / metabolism*
  • Intercellular Signaling Peptides and Proteins*
  • Iodine Radioisotopes / metabolism
  • Lung / metabolism
  • Mink
  • Progranulins
  • Receptors, Cell Surface / chemistry*
  • Recombinant Proteins / metabolism
  • Succinimides / metabolism

Substances

  • Affinity Labels
  • Cross-Linking Reagents
  • Glycoproteins
  • Growth Substances
  • Intercellular Signaling Peptides and Proteins
  • Iodine Radioisotopes
  • Progranulins
  • Receptors, Cell Surface
  • Recombinant Proteins
  • Succinimides
  • disuccinimidyl suberate