The 170-kDa glucose-regulated stress protein Is an endoplasmic reticulum protein that binds immunoglobulin

Hung Yun Lin, Patricia Masso-Welch, Yuan Pu Di, Jei Wei Cai, Jun Wen Shen, John R. Subjeck

Research output: Contribution to journalArticle

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Abstract

Anoxia, glucose starvation, calcium ionophore A23187, EDTA, glucosamine, and several other conditions that adversely affect the function of the endoplasmic reticulum (ER) induce the synthesis of the glucose-regulated class of stress proteins (GRPs). The primary GRPs induced by these stresses migrate at 78 and 94 kDa (GRP 78 and GRP94). In addition, another protein of ∼ 150-170 kDa (GRP 170) has been previously observed and is coordinately induced with GRP78 and GRP94. To characterize this novel stress protein, we have prepared an antisera against purified GRP170. Immunofluorescence, Endoglycosidase H sensitivity, and protease resistance of this protein in microsomes indicates that GRP170 is an ER lumenal glycoprotein retained in a pre-Golgi compartment. Immunoprecipitation of GRP170 with our antibody coprecipitates the GRP 78 (also referred to as the B cell immunoglobulin-binding protein) and GRP94 members of this stress protein family in Chinese hamster ovary cells under stress conditions. ATP depletion, by immunoprecipitation in the presence of apyrase, does not affect the interaction between GRP78 and GRP170 but results in the coprecipitation of an unidentified 60-kDa protein. In addition, GRP170 is found to be coprecipitated with immunoglobulin (Ig) in four different B cell hybridomas expressing surface IgM, cytoplasmic Ig light chain only, cytoplasmic Ig heavy chain only, or an antigen specific secreted IgG. In addition, in IgM surface expressing WEHI-231 B cells, anti-IgM coprecipitates GRP78, GRP94, as well as GRP170; antibodies against GRP170 and GRP94 reciprocally coprecipitate GRP94/GRP170 as well as GRP78. Results suggest that this 170-kDa GRP is a retained ER lumenal glycoprotein that is constitutively present and that may play a role in immunoglobulin folding and assembly in conjunction or consecutively with GRP78 and GRP94.

Original languageEnglish
Pages (from-to)1109-1119
Number of pages11
JournalMolecular Biology of the Cell
Volume4
Issue number11
DOIs
Publication statusPublished - Jan 1 1993
Externally publishedYes

Fingerprint

Endoplasmic Reticulum
Immunoglobulins
Heat-Shock Proteins
Proteins
Glucose
B-Lymphocytes
Immunoprecipitation
Immunoglobulin M
Glycoproteins
glucose-regulated protein 170
Apyrase
Immunoglobulin Light Chains
Immunoglobulin Heavy Chains
Calcium Ionophores
Antibodies
Glycoside Hydrolases
Glucosamine
Calcimycin
Hybridomas
Microsomes

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

Cite this

The 170-kDa glucose-regulated stress protein Is an endoplasmic reticulum protein that binds immunoglobulin. / Lin, Hung Yun; Masso-Welch, Patricia; Di, Yuan Pu; Cai, Jei Wei; Shen, Jun Wen; Subjeck, John R.

In: Molecular Biology of the Cell, Vol. 4, No. 11, 01.01.1993, p. 1109-1119.

Research output: Contribution to journalArticle

Lin, Hung Yun ; Masso-Welch, Patricia ; Di, Yuan Pu ; Cai, Jei Wei ; Shen, Jun Wen ; Subjeck, John R. / The 170-kDa glucose-regulated stress protein Is an endoplasmic reticulum protein that binds immunoglobulin. In: Molecular Biology of the Cell. 1993 ; Vol. 4, No. 11. pp. 1109-1119.
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