The distribution of mass in heteropolymer intermediate filaments assembled in vitro. Stem analysis of vimentin/desmin and bovine epidermal keratin.
We have studied the distribution of mass in several types of intermediate filaments (IF) assembled in vitro, by analyzing scanning transmission electron micrographs (STEM) of unstained specimens imaged in dark-field mode. Bovine epidermal keratin IF, which are obligate heteropolymers, were thus characterized and compared with facultative heteropolymers of vimentin and desmin and with earlier observations of homopolymer vimentin IF. The major components of each type of IF have linear densities of 37 kilodaltons/nm. Minor polymorphic variants are also present in each case, with linear densities of approximately 25 and 48 kilodaltons/nm, respectively. In view of the known subunit masses, these results are consistent with the proposition that at least three IF types (bovine epidermal keratin, vimentin, and desmin) are structurally homologous. For each type of IF studied, measurements taken near the ends of long IF or from short IF tend to have lower densities, characteristic of the least dense polymorphic variant. This form may represent a precursor "minimal core" structure in the sense of a minimal aggregate of protofilaments sufficiently stable to permit elongation. In the course of in vitro assembly, this form would be maturable to the normal IF structure at 37 kilodaltons/nm by the accretion of further protein. We also find the projected widths of these IF as measured from the STEM images to be significantly greater than the corresponding diameters determined in previous studies by conventional electron microscopy. Whereas the conventional techniques have routinely yielded diameters of 7-11 nm for IF contrasted by heavy metal staining, the STEM images of unstained IF provide estimates of 15 +/- 1 nm for the outer diameter of the major density class for each IF type studied and average values of 13.8 and 16.1 nm for the less dense and more dense variants, respectively.
D000818:Animals; D002417:Cattle; D003599:Cytoskeleton / Q000648:ultrastructure*; D003893:Desmin; D007381:Intermediate Filament Proteins / Q000032:analysis*; D007633:Keratins / Q000032:analysis*; D046911:Macromolecular Substances; D008854:Microscopy, Electron; D008855:Microscopy, Electron, Scanning; D008970:Molecular Weight; D012867:Skin / Q000032:analysis*; D014746:Vimentin
D003893:Desmin; D007381:Intermediate Filament Proteins; D046911:Macromolecular Substances; D014746:Vimentin; D007633:Keratins