There is no author summary for this article yet. Authors can add summaries to their articles on ScienceOpen to make them more accessible to a non-specialist audience.
Abstract
Two commercially available monoclonal antibodies raised against the intermediate filament
protein vimentin were characterized concerning their species-specific reaction pattern
on vertebrate cells. The antibody V9 exhibited extensive reactivity with vimentin
of all mammalian species tested, but specifically did not detect vimentin in mouse
cells and chicken fibroblasts. The antibody VIM 3B4 recognized vimentin in cells of
chicken and most mammalian species, except for rodent species. Characterization of
the binding site of VIM 3B4 on human vimentin by limited proteolysis and immunoblotting
as well as by sequence comparison strongly suggested that the epitope is located in
the coil 2 part of the vimentin rod domain. Site-directed mutagenesis of a mouse vimentin
cDNA clone followed by in vivo expression showed that VIM 3B4 could detect rodent
vimentin containing a single amino acid substitution (valine for leucine) at position
353 of the mouse vimentin sequence. Practical application for this finding was demonstrated
by the unequivocal identification of a modified murine vimentin protein, distinct
from the endogenous vimentin, in a cytoplasmic intermediate filament network in mouse
skin fibroblasts transfected with a recombinant plasmid expression vector.