Purification of human tumor cell autocrine motility factor and molecular cloning of its receptor

J Biol Chem. 1991 Jul 15;266(20):13442-8.

Abstract

Tumor autocrine motility factor (AMF) has been detected in and purified from serum-free conditioned medium of human HT-1080 fibrosarcoma cells. Under nonreducing conditions, AMF migrates in sodium dodecyl sulfate-polyacrylamide gel electrophoresis as a single band of 55 kDa but under reducing conditions as a band of 64 kDa. Two-dimensional polyacrylamide gel electrophoresis of the purified AMF resolved two groups of polypeptides with isoelectric points of 6.1 and 6.2 (majors), 6.35 and 6.4 (minors). Purified AMF stimulated HT-1080 cell migration in a dose-dependent fashion. The motility stimulation of the fibrosarcoma cells with AMF is associated with the phosphorylation of the AMF receptor, a 78-kDa cell surface glycoprotein (gp78), suggesting protein kinase participation in migratory signal transduction. The gene encoding gp78 was cloned from an HT-1080 fibrosarcoma complementary DNA library. The deduced sequence encodes a polypeptide of 323 amino acids. The nucleotide and predicted amino acid sequence of the gp78 reveals significant homology with the human suppressor/oncogene p53 protein.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Antibodies, Monoclonal
  • Base Sequence
  • Biomarkers, Tumor / isolation & purification*
  • Cell Line
  • Cell Movement / drug effects
  • Cloning, Molecular
  • DNA, Neoplasm / genetics
  • DNA, Neoplasm / isolation & purification
  • Fibrosarcoma
  • Gene Library
  • Glucose-6-Phosphate Isomerase
  • HeLa Cells / physiology
  • Humans
  • Molecular Sequence Data
  • Neoplasm Proteins / isolation & purification*
  • Neoplasm Proteins / metabolism
  • Neoplasm Proteins / pharmacology
  • RNA, Neoplasm / genetics
  • RNA, Neoplasm / isolation & purification
  • Receptors, Autocrine Motility Factor
  • Receptors, Cell Surface / genetics*
  • Receptors, Cytokine*
  • Sequence Homology, Nucleic Acid
  • Tumor Suppressor Protein p53 / genetics
  • Ubiquitin-Protein Ligases

Substances

  • Antibodies, Monoclonal
  • Biomarkers, Tumor
  • DNA, Neoplasm
  • Neoplasm Proteins
  • RNA, Neoplasm
  • Receptors, Cell Surface
  • Receptors, Cytokine
  • Tumor Suppressor Protein p53
  • AMFR protein, human
  • Receptors, Autocrine Motility Factor
  • Ubiquitin-Protein Ligases
  • Glucose-6-Phosphate Isomerase

Associated data

  • GENBANK/D10110
  • GENBANK/D10111
  • GENBANK/M63175
  • GENBANK/M63255
  • GENBANK/M63926
  • GENBANK/M63977
  • GENBANK/M63978
  • GENBANK/M63997
  • GENBANK/M63998
  • GENBANK/M64865