Extended Data Fig. 10: Testing the role of disulfide reduction in the degradation of CPY* and homologs of Mnl1. | Nature Structural & Molecular Biology

Extended Data Fig. 10: Testing the role of disulfide reduction in the degradation of CPY* and homologs of Mnl1.

From: Initiation of ERAD by the bifunctional complex of Mnl1/Htm1 mannosidase and protein disulfide isomerase

Extended Data Fig. 10

(a) ERAD of CPY*-HA was determined by cycloheximide (CHX) chase experiments in wild-type (WT) cells or cells lacking Gsh1 (gsh1Δ). The cells were grown in the presence of different concentrations of GSH before addition of CHX. The samples were analyzed by SDS-PAGE and immunoblotting for HA. Blotting for Pgk1 served as a loading control. Quantification of three similar experiments is shown in Fig. 7g. (b) As in (a), but following ERAD of CPY*-HA and a mutant lacking all cysteines (CPY*ΔCys) in the indicated cells grown in a low concentration of GSH (0.1 μM). Where indicated, Mnl1 was overexpressed from the GAL1 promoter (Mnl1 OE). Quantification of three similar experiments is shown in Fig. 7h. (c) Domain organizations of Schizosaccharomyces pombe Mnl1 (spMnl1) and of the mammalian homologs of Mnl1 (EDEMs).

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