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Related Experiment Videos

Murine antiestrogen-binding protein: characterization, solubilization and modulation by lipids.

A Matin1, P L Hwang, O L Kon

  • 1Department of Physiology, National University of Singapore, Kent Ridge.

Biochimica Et Biophysica Acta
|December 10, 1987
PubMed
Summary

Mouse antiestrogen-binding protein, found in liver endoplasmic reticulum, binds [3H]tamoxifen with high affinity. Its binding is inhibited by fatty acids and 7-ketocholesterol, with lysine residues potentially involved.

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Pharmacology

Background:

  • Antiestrogen-binding protein (AEBP) distribution and properties vary across species.
  • Nonsteroidal antiestrogens act as pure agonists in mice.
  • Understanding AEBP is crucial for antiestrogen drug development.

Purpose of the Study:

  • To characterize the antiestrogen-binding protein in mouse tissues.
  • To investigate the subcellular localization and binding characteristics of murine AEBP.
  • To elucidate the molecular properties of AEBP involved in antiestrogen binding.

Main Methods:

  • Tissue distribution analysis of AEBP in mice.
  • Subcellular fractionation of mouse liver.
  • Binding assays using [3H]tamoxifen.

Related Experiment Videos

  • Inhibition studies with fatty acids and 7-ketocholesterol.
  • Protein solubilization and characterization (molecular mass, sedimentation coefficient).
  • Proteolytic digestion to identify binding site residues.

    • Main Results:

    • AEBP is present in all mouse tissues, with highest concentrations in the liver.
    • In mouse liver, 82% of AEBP is localized to rough endoplasmic reticulum membranes.
    • AEBP exhibits high-affinity binding to [3H]tamoxifen (Kd = 1 nM).
    • Binding is inhibited by unsaturated fatty acids (cis-isomers more effective) and 7-ketocholesterol.
    • Solubilized AEBP has a molecular mass of ~700 kDa and a sedimentation coefficient of ~19 S.
    • Trypsin and non-specific proteinases abolish [3H]tamoxifen binding, suggesting lysine involvement.

    Conclusions:

    • Murine antiestrogen-binding protein is predominantly localized in the endoplasmic reticulum.
    • AEBP displays high-affinity binding to tamoxifen, modulated by lipids and cholesterol derivatives.
    • Lysine residues are likely critical for [3H]tamoxifen binding to AEBP.