Estrogen receptor preparation effects on the receptor-DNA interaction by surface plasmon resonance

Anal Bioanal Chem. 2016 Nov;408(29):8257-8262. doi: 10.1007/s00216-016-9967-0. Epub 2016 Oct 8.

Abstract

Up to now, several studies have investigated estrogen receptor (ER)-estrogen response element (ERE) interaction using biosensors such as surface plasmon resonance. These strategies have aimed to understand the molecular mechanism of such interaction as well as the effect of the ligand on this interaction. These approaches start to be used to determine the mechanisms of protein/DNA interaction, in particular in the context of drug discovery or environmental applications. However, some physical and biochemical parameters (incubation time, temperature, protease inhibitor cocktail, and bovine serum albumin (BSA)) are not completely described in the literature and could deeply modify the obtained results. This paper aims to focus not only on the preliminary steps of sample preparation such as protein thawing and incubation conditions (time and temperature) but also on the evaluation of protease inhibitor cocktail and BSA effect on the measurement of ER-ERE interactions.

Keywords: Dimerization; Estrogen; Estrogen receptor; Surface plasmon resonance; Temperature.

MeSH terms

  • Biosensing Techniques / methods*
  • DNA / chemistry*
  • Estradiol / chemistry*
  • Estrogen Receptor alpha / chemistry*
  • Humans
  • Protein Binding
  • Protein Denaturation
  • Protein Multimerization
  • Response Elements*
  • Serum Albumin, Bovine / chemistry
  • Surface Plasmon Resonance*

Substances

  • ESR1 protein, human
  • Estrogen Receptor alpha
  • Serum Albumin, Bovine
  • Estradiol
  • DNA