Subunit composition of the heteromeric cytosolic aryl hydrocarbon receptor complex

Huey Shang Chen, Gary H. Perdew

Research output: Contribution to journalArticle

126 Citations (Scopus)

Abstract

In a previous cross-linking study we have shown that the cytosolic aryl hydrocarbon receptor (AhR) complex has a heterotetrameric structure (Perdew, G. H. (1992) Biochem. Biophys. Res. Commun. 182, 55-62). In this report, both cross-linked and [35S]methionine-labeled Hepa 1c1c7 cytosol were used to characterize the subunit composition of the AhR complex by immunoprecipitation with an AhR polyclonal antibody followed by immunochemical analysis using antibodies against the AhR and 90-kDa heat shock protein (hsp90). Results indicated that the four subunits found in cross-linking experiments were composed of three species: the AhR ligand binding subunit hsp90, and an unknown 43-kDa protein. The stoichiometry of hsp90 present in each AhR complex was determined in two separate experiments: 1) from cross-linking experiments, stoichiometry was determined by quantitative immunoblotting with anti-AhR and anti-hsp90 antibodies followed by quantitation with 125I-counterantibody on protein blots; 2) using 35S-labeled Hepa 1 cytosol, the hsp90/AhR stoichiometry was determined by immunopurifying receptor complexes, and the amount of 35S-labeled AhR and hsp90 was assessed. The stoichiometry values obtained were 2.4 and 1.72 mol of hsp90/mol of AhR using each experimental approach, respectively.

Original languageEnglish (US)
Pages (from-to)27554-27558
Number of pages5
JournalJournal of Biological Chemistry
Volume269
Issue number44
StatePublished - Nov 4 1994

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Aryl Hydrocarbon Receptors
Chemical analysis
Stoichiometry
Cytosol
Antibodies
HSP90 Heat-Shock Proteins
Experiments
Heat-Shock Proteins
Immunoprecipitation
Immunoblotting
Methionine
Anti-Idiotypic Antibodies
Proteins
Ligands

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

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abstract = "In a previous cross-linking study we have shown that the cytosolic aryl hydrocarbon receptor (AhR) complex has a heterotetrameric structure (Perdew, G. H. (1992) Biochem. Biophys. Res. Commun. 182, 55-62). In this report, both cross-linked and [35S]methionine-labeled Hepa 1c1c7 cytosol were used to characterize the subunit composition of the AhR complex by immunoprecipitation with an AhR polyclonal antibody followed by immunochemical analysis using antibodies against the AhR and 90-kDa heat shock protein (hsp90). Results indicated that the four subunits found in cross-linking experiments were composed of three species: the AhR ligand binding subunit hsp90, and an unknown 43-kDa protein. The stoichiometry of hsp90 present in each AhR complex was determined in two separate experiments: 1) from cross-linking experiments, stoichiometry was determined by quantitative immunoblotting with anti-AhR and anti-hsp90 antibodies followed by quantitation with 125I-counterantibody on protein blots; 2) using 35S-labeled Hepa 1 cytosol, the hsp90/AhR stoichiometry was determined by immunopurifying receptor complexes, and the amount of 35S-labeled AhR and hsp90 was assessed. The stoichiometry values obtained were 2.4 and 1.72 mol of hsp90/mol of AhR using each experimental approach, respectively.",
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Subunit composition of the heteromeric cytosolic aryl hydrocarbon receptor complex. / Chen, Huey Shang; Perdew, Gary H.

In: Journal of Biological Chemistry, Vol. 269, No. 44, 04.11.1994, p. 27554-27558.

Research output: Contribution to journalArticle

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