Neurofilaments from ox spinal nerves. Isolation, disassembly, reassembly and cross-linking properties.
An isolation procedure for neurofilaments from ox spinal nerves is described where the triplet polypeptides (which have molecular weights of 205 000, 158 000 and 72 000) constitute more than 80% of the preparation. Soon after purification, the neurofilaments form a gel that is stable for many weeks. The purified neurofilaments disassemble in low-salt buffers at pH greater than 7.0 into soluble particles that contain all of the triplet polypeptides. Greater than 90% of the protein can reassemble to form filaments. The thiol-containing residues in the filaments can be cross-linked. Analyses of the complexes formed show that in the filament the 205 000-mol.wt. components are arranged to that they can be cross-linked to themselves and to the 158 000-mol.wt. polypeptides, and that the 72 000-mol.wt. components are arranged so that their thiol groups can be cross-linked together.
D000818:Animals; D002417:Cattle; D002458:Cell Fractionation / Q000379:methods; D003432:Cross-Linking Reagents / Q000494:pharmacology; D003599:Cytoskeleton / Q000032:analysis / Q000648:ultrastructure; D004220:Disulfides / Q000032:analysis; D004591:Electrophoresis, Polyacrylamide Gel; D007381:Intermediate Filament Proteins / Q000302:isolation & purification / Q000378:metabolism; D046911:Macromolecular Substances; D008297:Male; D008854:Microscopy, Electron; D008970:Molecular Weight; D016900:Neurofilament Proteins; D010618:Phenanthrolines / Q000494:pharmacology; D013127:Spinal Nerves / Q000032:analysis / Q000648:ultrastructure*
D003432:Cross-Linking Reagents; D004220:Disulfides; D007381:Intermediate Filament Proteins; D046911:Macromolecular Substances; D016900:Neurofilament Proteins; D010618:Phenanthrolines; C025205:1,10-phenanthroline