spacer gif spacer gif spacer gif spacer gif spacer gif
QUICK SEARCH:   [advanced]


spacer gif
     Home     Help     Feedback     Subscriptions     Archive     Search     Table of Contents    

Right arrow Help viewing high resolution images
Right arrow Return to article
(Downloading may take up to 30 seconds.
If the slide opens in your browser, select File -> Save As to save it.)
Click on image to view larger version.


Figure 8


Fig. 8. Interaction of DP with wild-type desmin and tailless desmin in GST pull-down experiments. (A) 35S-radiolabeled recombinant forms of wild-type desmin and tailless desmin were generated by coupled in vitro transcription/translation and analyzed by SDS-PAGE and autoradiography. Their Mr was 54K and 48K, respectively, as predicted on the basis of their cDNA. Ladder: molecular mass markers, Mr from top to bottom: 175K, 83K, 62K, 47.5K and 32.5K. (B) GST-fusion proteins of DP-BCS2849G and of BP230-BC were immobilized on glutathione agarose beads and incubated with either 35S-labeled wild-type desmin or tailless desmin. After washing, samples were resolved by SDS-PAGE and 35S-labeled bound proteins were visualized by autoradiography (C). Note that wild-type and tailless desmin showed a tendency to proteolytic degradation under the conditions of the GST pull-down experiments. (B) Coomassie blue staining of the gel depicted in panel B to verify equimolar loading of GST fusion proteins. (D) Different amounts of polymerized wild-type and tailless desmin were immobilized on a nitrocellulose membrane by dot blotting and incubated with radiolabeled DP-BC. Protein loading was verified by amino-black staining of the spotted proteins. The binding efficiency of DP-BC to both wild-type and tailless desmin was not significantly different and abruptly decreased when the amount of desmin was below 0.5'g. Note that binding of radiolabeled DP-BCS2849G to wild-type desmin was comparable with that of DP-BC.





Right arrow Return to article