Oligomerization of the tetramerization domain of p53 probed by two- and three-color single-molecule FRET.

ABSTRACT: We describe a method that combines two- and three-color single-molecule FRET spectroscopy with 2D FRET efficiency-lifetime analysis to probe the oligomerization process of intrinsically disordered proteins. This method is applied to the oligomerization of the tetramerization domain (TD) of the tumor suppressor protein p53. TD exists as a monomer at subnanomolar concentrations and forms a dimer and a tetramer at higher concentrations. Because the dissociation constants of the dimer and tetramer are very close, as we determine in this paper, it is not possible to characterize different oligomeric species by ensemble methods, especially the dimer that cannot be readily separated. However, by using single-molecule FRET spectroscopy that includes measurements of fluorescence lifetime and two- a

SUBMITTER: Chung HS 

PROVIDER: S-EPMC5565410 | biostudies-literature | 2017 Aug

REPOSITORIES: biostudies-literature