Identification of phosphorylation sites in the mouse oestrogen receptor

Lahooti, H., White, R., Hoare, S. A., Rahman, D., Pappin, D. J., Parker, M. G. (December 1995) Identification of phosphorylation sites in the mouse oestrogen receptor. J Steroid Biochem Mol Biol, 55 (3-4). pp. 305-13. ISSN 0960-0760 (Print)0960-0760 (Linking)

DOI: 10.1016/0960-0760(95)00188-3


Phosphorylation sites in the mouse oestrogen receptor, expressed in COS-1 cells in the presence of 17 beta-oestradiol, have been mapped by solid phase microsequencing. The receptor was first radio-labelled with [32P]orthophosphate and a number of 3H- or 14C-labelled amino acids, immunopurified and then tryptic peptides were separated by thin layer chromatography or high performance liquid chromatography. Amino acid sequence analysis indicated that Ser-122, Ser-156, Ser-158 and Ser-298 were phosphorylated. The substitution of Ser-122 and Ser-298 with alanine had a negligible effect on the transcriptional activity of the receptor in transfected cells. However, a reduction of transcriptional activity was observed when Ser-122 was mutated in the context of mutations in a putative amphipathic alpha-helix involved in AF-2 activity. Thus a region of AF-1 that encompasses Ser-122 appears to interact with AF-2 in the full-length receptor.

Item Type: Paper
Uncontrolled Keywords: 3T3 Cells Alanine Amino Acid Sequence Animals Base Sequence Binding Sites Cell Line/metabolism Chromatography, High Pressure Liquid Estradiol/metabolism Mice Molecular Sequence Data Mutation Peptide Fragments/chemistry Phosphorylation Phosphoserine Receptors, Estrogen/chemistry/ genetics/ metabolism Sequence Analysis/methods Transcription, Genetic Transfection
Subjects: bioinformatics > genomics and proteomics > design > amino acid design
organism description > animal > mammal > rodent > mouse
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein expression > phosphorylation
CSHL Authors:
Communities: CSHL labs > Pappin lab
Depositing User: Jessica Koos
Date: December 1995
Date Deposited: 08 Aug 2014 20:40
Last Modified: 08 Aug 2014 20:40
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