[Vimentin and Nup 180 in vitro binding assay].
In order to investigate relationship between vimentin and nuclear pore complex, we examined binding ability of vimentin, expressed in E. coli, with nucleoporin, isolated from rat liver nuclei, in vitro. Negative staining electron microscopy showed that the vimentin expressed in bacteria assembled 10 nm filament in vitro. SDS-PAGE and western blotting showed that Nup 180 bind to vimentin in binding assay in vitro. Combining immunogold labeling and negative staining electron microscopy techniques, we showed that Nup 180 bind on the 10 nm vimentin filaments. The experiment results indicated that vimentin filament may be anchored on nuclear pore complex in vivo by binding with Nup 180.
Cai
S T
ST
Institute of Biophysics, Academia Sinica, Beijing.
Zhou
F L
FL
Zhang
J Z
JZ
D000818:Animals; D001667:Binding, Competitive; D004926:Escherichia coli / Q000378:metabolism; D008099:Liver / Q000166:cytology / Q000378:metabolism; D028861:Nuclear Pore Complex Proteins; D009687:Nuclear Proteins / Q000302:isolation & purification / Q000378:metabolism*; D051381:Rats; D014746:Vimentin / Q000096:biosynthesis / Q000378:metabolism*
D028861:Nuclear Pore Complex Proteins; D009687:Nuclear Proteins; C513083:Nup153 protein, rat; D014746:Vimentin