Studies of interaction of tumor suppressor p53 with apo-MT using surface plasmon resonance

Anal Bioanal Chem. 2009 Dec;395(8):2569-75. doi: 10.1007/s00216-009-3174-1. Epub 2009 Oct 29.


The interaction of tumor suppressor p53 with apo-metallothionein (apo-MT) has been carried out using a flow injection-surface plasmon resonance (FI-SPR) instrument. MT was first tethered onto the carboxymethylated dextran film. Via incorporation of glycine-HCl (pH 2) to remove the sequestrated metal ions inherent in MT molecules, a more extended and open structure of apo-MT was formed. Substantial SPR angle shift corresponding to the interaction of wild-type p53 with apo-MT was observed. The interaction was originated from the binding between the free sulfhydryl groups of apo-MT and Zn(2+) of p53 with the binding constant of 1.4 x 10(8) M(-1). The specific binding of p53 to consensus double-stranded DNA was hindered after metal chelation from p53 by apo-MT. Furthermore, inhibition of the interaction between p53 and apo-MT imposed by p53/DNA complex was observed. The fluorescence measurements also revealed the binding of p53 to apo-MT, being consistent with the SPR results. Thus, SPR could potentially serve as an attractive technique for monitoring p53 conformational change and transcriptional activity regulated by the MT/apo-MT couple.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Biosensing Techniques
  • DNA / metabolism*
  • Humans
  • Liver / metabolism
  • Metallothionein / chemistry*
  • Metallothionein / metabolism*
  • Protein Binding
  • Rabbits
  • Spectrometry, Fluorescence
  • Surface Plasmon Resonance / methods*
  • Tumor Suppressor Protein p53 / chemistry*
  • Tumor Suppressor Protein p53 / metabolism*


  • Tumor Suppressor Protein p53
  • DNA
  • Metallothionein