Aspects of the metabolism of the epidermal growth factor receptor in A431 human epidermoid carcinoma cells

Mol Cell Biol. 1984 Apr;4(4):571-5. doi: 10.1128/mcb.4.4.571-575.1984.

Abstract

The biosynthesis and posttranslational metabolism of the epidermal growth factor (EGF) receptor were examined in the A431 human epidermoid carcinoma cell line. Polyclonal antibody against the receptor specifically immunoprecipitated two [35S]methionine-labeled proteins of Mr = 160,000 and 170,000. Pulse chase experiments showed the Mr = 160,000 protein to be a precursor of the Mr = 170,000 protein. Preincubation with tunicamycin resulted in immunoprecipitation of a single band of Mr = 130,000, whereas monensin inhibited maturation to the Mr = 170,000 form. Digestion of the Mr = 160,000 and 170,000 proteins with endoglycosidase H resulted in the appearance of Mr = 130,000 and 165,000 proteins, respectively. Prolonged pulse-chase experiments indicated that the half-life of the receptor is ca. 20 h in the absence of EGF and 5 h in the presence of EGF. Approximately three- to five-fold more phosphate is incorporated into the mature receptor upon addition of EGF, due primarily to increases in levels of phosphotyrosine and phosphoserine. Phosphate was also present on the Mr = 160,000 protein and the Mr = 130,000 protein found in the presence of tunicamycin.

Publication types

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

MeSH terms

  • Antibodies, Monoclonal
  • Carcinoma, Squamous Cell / metabolism*
  • Cell Line
  • Cells, Cultured
  • ErbB Receptors
  • Glycoproteins / biosynthesis
  • Humans
  • Molecular Weight
  • Phosphorylation
  • Protein Processing, Post-Translational
  • Receptors, Cell Surface / immunology
  • Receptors, Cell Surface / metabolism*

Substances

  • Antibodies, Monoclonal
  • Glycoproteins
  • Receptors, Cell Surface
  • ErbB Receptors