Purification and Structural Analysis of Desmoplakin

Methods Enzymol. 2016:569:197-213. doi: 10.1016/bs.mie.2015.05.006. Epub 2015 May 30.

Abstract

Desmoplakin (DP) is an obligate component of desmosomes, where it links the desmosomal cadherin/plakoglobin/plakophilin assembly to intermediate filaments. DP contains a large amino-terminal domain (DPNT) that binds to the cadherin/plakoglobin/plakophilin complex, a central coiled-coil domain that dimerizes the molecule, and a C-terminal domain (DPCT) that binds to intermediate filaments. DPNT contains a plakin domain, comprising a set of spectrin-like repeats. DPCT contains three plakin repeat domains, each formed by 4.5 repeats of a sequence motif known as a plakin repeat that bind to intermediate filaments. Here, we review purification, biochemical characterization, and structural analysis of the DPNT plakin domain and the DPCT plakin repeat domains.

Keywords: Desmoplakin; Desmosome; Intermediate filament; Plakin domain; Plakin repeat; Plakoglobin; Plakophilin; Spectrin repeat; Vimentin.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Chromatography, Gel
  • Chromatography, Ion Exchange
  • Desmoplakins / chemistry
  • Desmoplakins / isolation & purification*
  • Escherichia coli
  • Humans
  • Models, Molecular
  • Protein Binding
  • Protein Structure, Secondary
  • Scattering, Small Angle
  • X-Ray Diffraction

Substances

  • DSP protein, human
  • Desmoplakins