Figure 4: CAND1 displaces F-box proteins from Cul1p.
From: CAND1 controls in vivo dynamics of the cullin 1-RING ubiquitin ligase repertoire
(a) Cul1p- and Pof10p-associated protein complexes were immunopurified from Δknd1 cells and incubated with 1 μg of recombinant His-tagged human CAND1 for 30 min. One microgram of bovine serum albumin was used as control instead of CAND1. The complexes were analysed for the levels of Cul1p and Pof10p-Myc by immunoblotting with Cul1p and Myc antibodies, and signals were quantified. The asterisk denotes a band present in the CAND1 preparation that weakly crossreacts with the α-Cul1p antiserum (possibly insect cell CUL1). (b) Pof10p-Myc complexes were isolated from Δknd1 and Δknd1 csn5 cells as in (a) and incubated for the indicated times with 1 μg BSA or CAND1 as indicated. The complexes were analysed for the levels of Cul1p and Pof10p-Myc by immunoblotting with Cul1p and Myc antibodies, and signals were quantified.
