title: The three-dimensional structure of trichocyte (hard alpha-) keratin intermediate filaments: features of the molecular packing deduced from the sites of induced crosslinks.
abstract: The spatial distribution of the crosslinks that can be induced between lysine residues in trichocyte (alpha-) keratin intermediate filaments (IF) using disulfosuccinimidyl tartrate has been analyzed in detail and the results used to provide information about the three-dimensional (3-D) structure. The pattern of inter-molecular interactions derived from earlier studies is essentially two-dimensional in that it involves projection on to a cylinder followed by unwrapping to give a sheet. Crosslinks are observed between molecular strands four apart and it is shown that this can only occur if the paths of the molecular strands through the IF are systematically distorted. These crosslinks are clustered axially at intervals of around 15 nm, a value closely related to the pitch length of the constituent coiled-coil molecules in the rod domains. The number of crosslinks between adjacent molecular strands shows a striking difference depending on lateral direction and provides support for the concept of a head-to-tail stacking of tetramers defined by the A(CN) mode of packing to form protofilament substructures in the fully formed IF. Each protofilament would consist of a pair of oppositely directed molecular strands stabilized by A(11) and A(22) interactions identified in earlier work. A detailed model for the IF in the reduced state comprising a ring of eight protofilaments is suggested. When combined with earlier studies of crosslink formation in the oxidized state, the present findings lead to the conclusion that there is a major reorganization of the molecular packing within the protofilaments during keratinization in vivo. Taken in conjunction with existing X-ray data on the fully keratinized structures, the new evidence for a protofilament substructure also enables a detailed 3-D model for the mature IF to be suggested.
authors.affiliation: 28 Satinay Drive, Noosa Parklands, Tewantin, Qld 4565, Australia.
authors.forename: R D B
authors.initials: RD
authors.lastname: Fraser
authors.forename: David A D
authors.initials: DA
authors.lastname: Parry
mesh_terms: D000818:Animals; D003432:Cross-Linking Reagents; D005583:Fourier Analysis; D021621:Imaging, Three-Dimensional; D007382:Intermediate Filaments / Q000648:ultrastructure*; D007633:Keratins / Q000737:chemistry / Q000648:ultrastructure*; D008239:Lysine / Q000737:chemistry; D008961:Models, Structural; D010084:Oxidation-Reduction; D017434:Protein Structure, Tertiary; D013388:Succinimides; D014961:X-Ray Diffraction
chemical_list: D003432:Cross-Linking Reagents; D013388:Succinimides; C018052:disuccinimidyl tartarate; D007633:Keratins; D008239:Lysine