title: Mechanism of intermediate filament recognition by plakin repeat domains revealed by envoplakin targeting of vimentin.
abstract: Plakin proteins form critical connections between cell junctions and the cytoskeleton; their disruption within epithelial and cardiac muscle cells cause skin-blistering diseases and cardiomyopathies. Envoplakin has a single plakin repeat domain (PRD) which recognizes intermediate filaments through an unresolved mechanism. Herein we report the crystal structure of envoplakin's complete PRD fold, revealing binding determinants within its electropositive binding groove. Four of its five internal repeats recognize negatively charged patches within vimentin via five basic determinants that are identified by nuclear magnetic resonance spectroscopy. Mutations of the Lys1901 or Arg1914 binding determinants delocalize heterodimeric envoplakin from intracellular vimentin and keratin filaments in cultured cells. Recognition of vimentin is abolished when its residues Asp112 or Asp119 are mutated. The latter slot intermediate filament rods into basic PRD domain grooves through electrosteric complementarity in a widely applicable mechanism. Together this reveals how plakin family members form dynamic linkages with cytoskeletal frameworks.
authors.affiliation: School of Cancer Sciences, University of Birmingham, Birmingham B15 2TT, UK.
authors.forename: Claudia
authors.initials: C
authors.lastname: Fogl
authors.affiliation: Institute of Immunology and Immunotherapy, University of Birmingham, Birmingham B15 2TT, UK.
authors.forename: Fiyaz
authors.initials: F
authors.lastname: Mohammed
authors.affiliation: School of Cancer Sciences, University of Birmingham, Birmingham B15 2TT, UK.
authors.forename: Caezar
authors.initials: C
authors.lastname: Al-Jassar
authors.affiliation: School of Cancer Sciences, University of Birmingham, Birmingham B15 2TT, UK.
authors.forename: Mark
authors.initials: M
authors.lastname: Jeeves
authors.affiliation: School of Cancer Sciences, University of Birmingham, Birmingham B15 2TT, UK.
authors.forename: Timothy J
authors.initials: TJ
authors.lastname: Knowles
authors.affiliation: Nanoscale Physics Research Laboratory, School of Physics and Astronomy, University of Birmingham, Birmingham B15 2TT, UK.
authors.forename: Penelope
authors.initials: P
authors.lastname: Rodriguez-Zamora
authors.affiliation: School of Biosciences, University of Birmingham, Birmingham B15 2TT, UK.
authors.forename: Scott A
authors.initials: SA
authors.lastname: White
authors.affiliation: Institute of Cancer and Genomic Sciences, University of Birmingham, Birmingham B15 2TT, UK.
authors.forename: Elena
authors.initials: E
authors.lastname: Odintsova
authors.affiliation: School of Cancer Sciences, University of Birmingham, Birmingham B15 2TT, UK.
authors.forename: Michael
authors.initials: M
authors.lastname: Overduin
authors.affiliation: School of Cancer Sciences, University of Birmingham, Birmingham B15 2TT, UK.
authors.forename: Martyn
authors.initials: M
authors.lastname: Chidgey
mesh_terms: D020816:Amino Acid Motifs; D000818:Animals; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000378:metabolism; D006801:Humans; D007382:Intermediate Filaments / Q000737:chemistry / Q000378:metabolism*; D008565:Membrane Proteins / Q000737:chemistry / Q000378:metabolism*; D008958:Models, Molecular; D010802:Phylogeny; D011485:Protein Binding; D011487:Protein Conformation; D011498:Protein Precursors / Q000737:chemistry / Q000378:metabolism*; D014746:Vimentin / Q000737:chemistry / Q000378:metabolism*
chemical_list: D008565:Membrane Proteins; D011498:Protein Precursors; D014746:Vimentin; C100754:envoplakin