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.



Fig. 6. Actin polymerization regulates the interaction between cortactin and dynamin. (A) Purified GST-Dyn-PRD (30 nM) was mixed with immobilized His-cortactin ranging from 0-200 nM in actin polymerization buffer plus G-actin (2 µM) and Arp2/3 complex (200 nM), and incubated at room temperature for 30 minutes. As a control, the reaction was also performed in the same buffer without G-actin and Arp2/3 complex. The samples were briefly centrifuged and the supernatants were analyzed for the presence of remaining GST-Dyn-PRD by immunoblotting with GST antibody. Amounts of GST-Dyn-PRD on the blot were quantified by digital scanning and normalized to the percentage of depletion. The resulting data were used to fit a rectangular hyperbola, yielding apparent Kd values as indicated. Inset, showing that GST has little binding activity to cortactin as compared to GST-Dyn-PRD. (B) The interaction of GST-Dyn-PRD and cortactin was carried out with G-actin plus Arp2/3 complex or G-actin without Arp2/3 complex. Cortactin was also preincubated with G-actin and Arp2/3 complex for 20 minutes prior to incubation with GST-Dyn-PRD for additional 20 minutes.





Right arrow Return to article