FIGURE 7.

The interaction site on ASC PYD for self-association overlaps with the interaction site for pyrin PYD. A, in vitro binding of purified soluble ASC PYD to bead-bound GST-ASC PYD in the absence or presence of purified pyrin PYD. Bound ASC PYD was detected using an anti-ASC antibody, and bound pyrin PYD was detected using an anti-His antibody. GST-ASC PYD was detected using Ponceau S stain. An amount representing 20% of the input of ASC PYD and pyrin PYD detected by Coomassie stain is shown. B, binding of in vitro translated [³⁵S]methionine-labeled pyrin to bead-bound GST-ASC PYD or GST in the absence or presence of purified soluble ASC PYD. Bound [³⁵S]methionine-labeled pyrin was detected by phosphorimaging, whereas GST, GST-ASC PYD, and soluble ASC PYD were detected using Coomassie stain. An amount representing 10% of the input of ³⁵S-labeled pyrin is shown.