title: A 'hot-spot' mutation alters the mechanical properties of keratin filament networks.
abstract: Keratins 5 and 14 polymerize to form the intermediate filament network in the progenitor basal cells of many stratified epithelia including epidermis, where it provides crucial mechanical support. Inherited mutations in K5 or K14 result in epidermolysis bullosa simplex (EBS), a skin-fragility disorder. The impact that such mutations exert on the intrinsic mechanical properties of K5/K14 filaments is unknown. Here we show, by using differential interference contrast microscopy, that a 'hot-spot' mutation in K14 greatly reduces the ability of reconstituted mutant filaments to bundle under crosslinking conditions. Rheological assays measure similar small-deformation mechanical responses for crosslinked solutions of wild-type and mutant keratins. The mutation, however, markedly reduces the resilience of crosslinked networks against large deformations. Single-particle tracking, which probes the local organization of filament networks, shows that the mutant polymer exhibits highly heterogeneous structures compared to those of wild-type filaments. Our results indicate that the fragility of epithelial cells expressing mutant keratin may result from an impaired ability of keratin polymers to be crosslinked into a functional network.
authors.affiliation: Department of Biological Chemistry, The Johns Hopkins University School of Medicine, Baltimore, Maryland 21205, USA.
authors.forename: L
authors.initials: L
authors.lastname: Ma
authors.forename: S
authors.initials: S
authors.lastname: Yamada
authors.forename: D
authors.initials: D
authors.lastname: Wirtz
authors.forename: P A
authors.initials: PA
authors.lastname: Coulombe
mesh_terms: D055592:Biophysical Phenomena; D001703:Biophysics; D003599:Cytoskeleton / Q000737:chemistry*; D004817:Epidermis / Q000737:chemistry*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007633:Keratins / Q000737:chemistry* / Q000235:genetics* / Q000648:ultrastructure; D008854:Microscopy, Electron; D009154:Mutation; D020125:Mutation, Missense; D010957:Plasmids / Q000378:metabolism; D011108:Polymers / Q000737:chemistry; D011994:Recombinant Proteins / Q000737:chemistry
chemical_list: D011108:Polymers; D011994:Recombinant Proteins; D007633:Keratins