Release of low density lipoprotein receptors from human fibroblasts or Hela cells by tryptic digestion

Nicola Di Ferrante, Patricia V. Donnelly, Daniela T. Di Ferrante, Salvatore Toma, Antonio Gotto

Research output: Contribution to journalArticle

4 Scopus citations

Abstract

Trypsin treatment of cultured normal human skin fibroblasts or Hela cells releases material which is retained on a low density lipoprotein (LDL)-Sepharose affinity column, may be eluted from it with 2.5 M KI and, after dialysis, agglutinates LDL or apo-B-coated formocells. Such agglutination is prevented by preincubation of the receptor with LDL in solution or with arginine-rich protamine. Trypsin treatment of "receptor defective" or "receptor negative" mutant fibroblasts releases material which is retained on LDL-Sepharose column but fails to agglutinate LDL-coated formocells. The receptor may be labeled with 6-[3H]-glucosamine·HCl and [3H]-leucine, it is inactivated by heating at 80°C for 10 min and may be obtained from normal fibroblasts or Hela cells, whether they were cultured in presence or in absence of lipoprotein-containing fetal calf serum.

Original languageEnglish (US)
Pages (from-to)366-373
Number of pages8
JournalBiochemical and Biophysical Research Communications
Volume84
Issue number2
DOIs
StatePublished - Sep 29 1978

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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