Conformational detection of p53's oligomeric state by FlAsH Fluorescence

Tawnya M. Webber, Andrew C. Allen, Wai Kit Ma, Rhett G. Molloy, Charisse N. Kettelkamp, Caitlin A. Dow, Matthew J Gage

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

The p53 tumor suppressor protein is a critical checkpoint in prevention of tumor formation, and the function of p53 is dependent on proper formation of the active tetramer. In vitro studies have shown that p53 binds DNA most efficiently as a tetramer, though inactive p53 is predicted to be monomeric in vivo. We demonstrate that FlAsH binding can be used to distinguish between oligomeric states of p53, providing a potential tool to explore p53 oligomerization in vivo. The FlAsH tetra-cysteine binding motif has been incorporated along the dimer and tetramer interfaces in the p53 tetramerization domain to create reporters for the dimeric and tetrameric states of p53, though the geometry of the four cysteines is critical for efficient FlAsH binding. Furthermore, we demonstrate that FlAsH binding can be used to monitor tetramer formation in real-time. These results demonstrate the potential for using FlAsH fluorescence to monitor protein-protein interactions in vivo.

Original languageEnglish (US)
Pages (from-to)66-70
Number of pages5
JournalBiochemical and Biophysical Research Communications
Volume384
Issue number1
DOIs
StatePublished - Jun 19 2009

Fingerprint

Cysteine
Fluorescence
Tumor Suppressor Protein p53
Oligomerization
Dimers
Tumors
Proteins
Geometry
DNA
Neoplasms
In Vitro Techniques

Keywords

  • FlAsH
  • Fluorescence
  • Oligomerization
  • p53
  • Tetramerization

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Cell Biology
  • Molecular Biology

Cite this

Conformational detection of p53's oligomeric state by FlAsH Fluorescence. / Webber, Tawnya M.; Allen, Andrew C.; Ma, Wai Kit; Molloy, Rhett G.; Kettelkamp, Charisse N.; Dow, Caitlin A.; Gage, Matthew J.

In: Biochemical and Biophysical Research Communications, Vol. 384, No. 1, 19.06.2009, p. 66-70.

Research output: Contribution to journalArticle

Webber, Tawnya M. ; Allen, Andrew C. ; Ma, Wai Kit ; Molloy, Rhett G. ; Kettelkamp, Charisse N. ; Dow, Caitlin A. ; Gage, Matthew J. / Conformational detection of p53's oligomeric state by FlAsH Fluorescence. In: Biochemical and Biophysical Research Communications. 2009 ; Vol. 384, No. 1. pp. 66-70.
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