RGD Reference Report - Recombinant expression of aryl hydrocarbon receptor for quantitative ligand-binding analysis. - Rat Genome Database

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Recombinant expression of aryl hydrocarbon receptor for quantitative ligand-binding analysis.

Authors: Fan, MQ  Bell, AR  Bell, DR  Clode, S  Fernandes, A  Foster, PM  Fry, JR  Jiang, T  Loizou, G  MacNicoll, A  Miller, BG  Rose, M  Shaikh-Omar, O  Tran, L  White, S 
Citation: Fan MQ, etal., Anal Biochem. 2009 Jan 15;384(2):279-87. Epub 2008 Oct 7.
RGD ID: 2325669
Pubmed: PMID:18938125   (View Abstract at PubMed)
PMCID: PMC2621304   (View Article at PubMed Central)
DOI: DOI:10.1016/j.ab.2008.10.003   (Journal Full-text)

Recombinant expression of the aryl hydrocarbon receptor (AhR) yields small amounts of ligand-binding-competent AhR. Therefore, Spodoptera frugiperda (Sf9) cells and baculovirus have been evaluated for high-level and functional expression of AhR. Rat and human AhR were expressed as soluble protein in significant amounts. Expression of ligand-binding-competent AhR was sensitive to the protein concentration of Sf9 extract, and coexpression of the chaperone p23 failed to affect the yield of functional ligand-binding AhR. The expression system yielded high levels of functional protein, with the ligand-binding capacity (Bmax) typically 20-fold higher than that obtained with rat liver cytosol. Quantitative estimates of the ligand-binding affinity of human and rat AhR were obtained; the Kd for recombinant rat AhR was indistinguishable from that of native rat AhR, thereby validating the expression system as a faithful model for native AhR. The human AhR bound TCDD with significantly lower affinity than the rat AhR. These findings demonstrate high-level expression of ligand-binding-competent AhR, and sufficient AhR for quantitative analysis of ligand binding.



Gene Ontology Annotations    Click to see Annotation Detail View

Molecular Function

  
Object SymbolSpeciesTermQualifierEvidenceWithNotesSourceOriginal Reference(s)
AhrRattoxic substance binding  IDA  RGD 

Objects Annotated

Genes (Rattus norvegicus)
Ahr  (aryl hydrocarbon receptor)


Additional Information