Figure 7.

Influence of clustering and phosphorylation on α-appendage interactions. (A) Free appendages versus clustered appendages have different ligand preferences. As an approximation to free appendages, we have incubated GST-appendages in brain extract and subsequently captured these by pouring the extract over a filter with GSH beads on top. This was compared with GST-appendages previously bound to beads and then incubated in brain extract. The Coomassie gel shows the major changes taking place and some proteins have been blotted. The identities of the Coomassie bands have not been determined by mass spectrometry. (B) α-Appendages incubated in phosphorylated, dephosphorylated and untreated brain extracts change their ligand preferences. The identities of dominant proteins by LC-MS/MS are given beside the Coomassie gel.