A potential role for the COOH-terminal domain in the lateral packing of type III intermediate filaments.
To identify sites of self-association in type III intermediate filament (IF) proteins, we have taken an "anti-idiotypic antibody" approach. A mAb (anti-Ct), recognizing a similar feature near the end of the rod domain of vimentin, desmin, and peripherin (epsilon site or epsilon epitope), was characterized. Anti-idiotypic antibodies, generated by immunizing rabbits with purified anti-Ct, recognize a site (presumably "complementary" to the epsilon epitope) common among vimentin, desmin, and peripherin (beta site or beta epitope). The beta epitope is represented in a synthetic peptide (PII) modeled after the 30 COOH-terminal residues of peripherin, as seen by comparative immunoblotting assays. Consistent with the idea of an association between the epsilon and the beta site, PII binds in vitro to intact IF proteins and fragments containing the epsilon epitope, but not to IF proteins that do not react with anti-Ct. Microinjection experiments conducted in vivo and filament reconstitution assays carried out in vitro further demonstrate that "uncoupling" of this site-specific association (by competition with PII or anti-Ct) interferes with normal IF architecture, resulting in the formation of filaments and filament bundles with diameters much greater than that of the normal IFs. These thick fibers are very similar to the ones observed previously when a derivative of desmin missing 27 COOH-terminal residues was assembled in vitro (Kaufmann, E., K. Weber, and N. Geisler. 1985. J. Mol. Biol. 185:733-742). As a molecular explanation, we propose here that the epsilon and the beta sites of type III IF proteins are "complementary" and associate during filament assembly. As a result of this association, we further postulate the formation of a surface-exposed "loop" or "hairpin" structure that may sterically prevent inappropriate filament-filament aggregation and regulate filament thickness.
Kouklis
P D
PD
Programme of Cell Biology, European Molecular Biology Laboratory, Heidelberg, Germany.
Papamarcaki
T
T
Merdes
A
A
Georgatos
S D
SD
D000818:Animals; D000888:Antibodies, Anti-Idiotypic; D000911:Antibodies, Monoclonal; D001666:Binding Sites, Antibody; D002460:Cell Line; D003893:Desmin / Q000378:metabolism; D000939:Epitopes / Q000032:analysis; D015151:Immunoblotting; D007381:Intermediate Filament Proteins / Q000276:immunology / Q000378:metabolism* / Q000648:ultrastructure; D008562:Membrane Glycoproteins; D051379:Mice; D016253:Microscopy, Immunoelectron; D009419:Nerve Tissue Proteins; D009479:Neuropeptides / Q000378:metabolism; D064531:Peripherins; D014746:Vimentin / Q000378:metabolism
D000888:Antibodies, Anti-Idiotypic; D000911:Antibodies, Monoclonal; D003893:Desmin; D000939:Epitopes; D007381:Intermediate Filament Proteins; D008562:Membrane Glycoproteins; D009419:Nerve Tissue Proteins; D009479:Neuropeptides; D064531:Peripherins; D014746:Vimentin