Cell surface localization of the 78 kD glucose regulated protein (GRP 78) induced by thapsigargin

Andrea Delpino, Pierluca Piselli, Daniela Vismara, Silvia Vendetti, Vittorio Colizzi

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Abstract

In the present study it was found that the synthesis of the 78 kD glucose-regulated protein (GRP 78 or BIP) is vigorously induced in human rabdomiosarcoma cells (TE 671/RD) following both short-term (1 h) and prolonged (18 h) exposure to 100 nM thapsigargin (Tg). Flow cytometric analysis with a specific anti-GRP 78 polyclonal antibody showed that Tg-treated cells express the GRP 78 on the plasma membrane. Cell surface localization of the Tg-induced GRP 78 was confirmed by biotinylation of membrane-exposed proteins and subsequent isolation of the biotin-labelled proteins by streptavidin/agarose affinity chromatography. It was found that a fraction of the Tg-induced GRP 78 is present among the biotin-labelled, surface-exposed, proteins, Conversely, the GRP 78 immunoprecipitated from unfractionated lysates of Tg-treated and biotin-reacted cells was found to be biotinylated. This is the first report demonstrating surface expression of GRP 78 in cells exposed to a specific GRP 78-inducing stimulus.

Original languageEnglish
Pages (from-to)21-26
Number of pages6
JournalMolecular Membrane Biology
Volume15
Issue number1
Publication statusPublished - Jan 1998

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Thapsigargin
Biotin
Membrane Proteins
Biotinylation
Agarose Chromatography
Affinity Chromatography
Cell Membrane
glucose-regulated proteins
Antibodies
Proteins

Keywords

  • Cell membrane
  • Grp 78
  • Thapsigargin

ASJC Scopus subject areas

  • Cell Biology

Cite this

Cell surface localization of the 78 kD glucose regulated protein (GRP 78) induced by thapsigargin. / Delpino, Andrea; Piselli, Pierluca; Vismara, Daniela; Vendetti, Silvia; Colizzi, Vittorio.

In: Molecular Membrane Biology, Vol. 15, No. 1, 01.1998, p. 21-26.

Research output: Contribution to journalArticle

Delpino, Andrea ; Piselli, Pierluca ; Vismara, Daniela ; Vendetti, Silvia ; Colizzi, Vittorio. / Cell surface localization of the 78 kD glucose regulated protein (GRP 78) induced by thapsigargin. In: Molecular Membrane Biology. 1998 ; Vol. 15, No. 1. pp. 21-26.
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