Binding of SGTA to Rpn13 selectively modulates protein quality control

Leznicki, Pawel; Korac-Prlic, Jelena; Kliza, Katarzyna; Husnjak, Koraljka; Nyathi, Yvonne; Dikic, Ivan; High, Stephen

Anti-Rpn13 and anti-20S proteasome antibodies were obtained from Enzo Life Sciences, anti-FLAG M2 antibody from Sigma, anti-Myc antibody (clone 4A6) from Upstate and anti-His tag antibody from Novagen. Mouse anti-SGTA antibody (clone 47-B) was purchased from Santa Cruz Biotechnology, whereas a chicken anti-SGTA antibody was made to order. Mouse anti-tubulin antibody was a gift from Keith Gull (University of Oxford, UK). Rabbit anti-tubulin, rabbit anti-GFP, rabbit anti-PSMD1 and chicken anti-Bag6 antibodies were from Abcam, whereas mouse anti-V5 antibody was purchased from Abcam and Serotec. The monoclonal anti-opsin tag antibody has been previously described (Leznicki et al., 2010). The HEK293 cell line stably expressing the hRpn11-HTBH plasmid (HEK293Rpn11-HTBH) was a kind gift of Lan Huang (University of California, Irvine, CA) (Wang et al., 2007). Stable, tetracycline-inducible HeLa Flp-In T-REx cells used to generate a stable line expressing OP91 were from Stephen Taylor (University of Manchester, UK) (Tighe et al., 2008; Wunderley et al., 2014).