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 language | English |
|---|---|
| Pages (from-to) | 21-26 |
| Number of pages | 6 |
| Journal | Molecular Membrane Biology |
| Volume | 15 |
| Issue number | 1 |
| Publication status | Published - Jan 1998 |
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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 journal › Article
}
TY - JOUR
T1 - Cell surface localization of the 78 kD glucose regulated protein (GRP 78) induced by thapsigargin
AU - Delpino, Andrea
AU - Piselli, Pierluca
AU - Vismara, Daniela
AU - Vendetti, Silvia
AU - Colizzi, Vittorio
PY - 1998/1
Y1 - 1998/1
N2 - 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.
AB - 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.
KW - Cell membrane
KW - Grp 78
KW - Thapsigargin
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M3 - Article
VL - 15
SP - 21
EP - 26
JO - Molecular Membrane Biology
JF - Molecular Membrane Biology
SN - 0968-7688
IS - 1
ER -