title: The rod domain of NF-L determines neurofilament architecture, whereas the end domains specify filament assembly and network formation.
abstract: Neurofilaments, assembled from NF-L, NF-M, and NF-H subunits, are the most abundant structural elements in myelinated axons. Although all three subunits contain a central, alpha-helical rod domain thought to mediate filament assembly, only NF-L self-assembles into 10-nm filaments in vitro. To explore the roles of the central rod, the NH2-terminal head and the COOH-terminal tail domain in filament assembly, full-length, headless, tailless, and rod only fragments of mouse NF-L were expressed in bacteria, purified, and their structure and assembly properties examined by conventional and scanning transmission electron microscopy (TEM and STEM). These experiments revealed that in vitro assembly of NF-L into bona fide 10-nm filaments requires both end domains: whereas the NH2-terminal head domain promotes lateral association of protofilaments into protofibrils and ultimately 10-nm filaments, the COOH-terminal tail domain controls lateral assembly of protofilaments so that it terminates at the 10-nm filament level. Hence, the two end domains of NF-L have antagonistic effects on the lateral association of protofilaments into higher-order structures, with the effect of the COOH-terminal tail domain being dominant over that of the NH2-terminal head domain. Consideration of the 21-nm axial beading commonly observed with 10-nm filaments, the approximate 21-nm axial periodicity measured on paracrystals, and recent cross-linking data combine to support a molecular model for intermediate filament architecture in which the 44-46-nm long dimer rods overlap by 1-3-nm head-to-tail, whereas laterally they align antiparallel both unstaggered and approximately half-staggered.
authors.affiliation: M.E. Müller Institute at the Biocenter, University of Basel, Switzerland.
authors.forename: S
authors.initials: S
authors.lastname: Heins
authors.forename: P C
authors.initials: PC
authors.lastname: Wong
authors.forename: S
authors.initials: S
authors.lastname: Müller
authors.forename: K
authors.initials: K
authors.lastname: Goldie
authors.forename: D W
authors.initials: DW
authors.lastname: Cleveland
authors.forename: U
authors.initials: U
authors.lastname: Aebi
mesh_terms: D000818:Animals; D002417:Cattle; D066298:In Vitro Techniques; D007382:Intermediate Filaments / Q000737:chemistry / Q000648:ultrastructure*; D046911:Macromolecular Substances; D051379:Mice; D008854:Microscopy, Electron; D016900:Neurofilament Proteins / Q000737:chemistry*; D011485:Protein Binding; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins; D013329:Structure-Activity Relationship
chemical_list: D046911:Macromolecular Substances; D016900:Neurofilament Proteins; D011994:Recombinant Proteins