Endosome and Golgi‐associated degradation (EGAD) of membrane proteins regulates sphingolipid metabolism See also Fig EV2 and Appendix Tables S4 and S5.

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Endosome and Golgi‐associated degradation (EGAD) of membrane proteins regulates sphingolipid metabolism See also Fig EV2 and Appendix Tables S4 and S5. AVacuolar proteolysis‐deficient pep4∆ and tul1∆ pep4∆ cells were labeled to saturation with heavy 13C615N2‐L‐lysine and then chased for 3 h in the presence of light L‐lysine, following quantitative proteome analysis by LC‐MS (two biological replicates). To analyze Dsc complex‐dependent protein turnover, the ratio of H/L ratios of tul1∆ pep4∆ over pep4∆ cells of 2511 proteins quantified in both strains was plotted, and proteins with a ratio of H/L ratios > 2 were selected. Red squares: regulated proteins with at least one predicted transmembrane (TM) domain; black triangles: regulated proteins without TM domains. BSchematic representation of the Dsc ubiquitin ligase complex (a top hit from the SGA screen) and the SPOTS complex (a putative Dsc substrate). SPT, serine palmitoyltransferase; LCB, long‐chain bases; SL, sphingolipids. C–FSDS–PAGE and Western blot analysis with the indicated antibodies (C, D) of total yeast lysates from WT cells and the indicated mutants; (E) of input and elution (with FLAG peptide) from denaturing FLAG‐Orm2 immunoprecipitations (IP) from WT cells and the indicated mutants. Control cells expressed untagged Orm2; (F) of input and elution (with FLAG peptide) from native FLAG‐Orm2 co‐immunoprecipitations from WT cells and the indicated mutants. The control strain expresses untagged Orm2. IF THIS IMAGE HAS BEEN PROVIDED BY OR IS OWNED BY A THIRD PARTY, AS INDICATED IN THE CAPTION LINE, THEN FURTHER PERMISSION MAY BE NEEDED BEFORE ANY FURTHER USE. PLEASE CONTACT WILEY'S PERMISSIONS DEPARTMENT ON PERMISSIONS@WILEY.COM OR USE THE RIGHTSLINK SERVICE BY CLICKING ON THE 'REQUEST PERMISSIONS' LINK ACCOMPANYING THIS ARTICLE. WILEY OR AUTHOR OWNED IMAGES MAY BE USED FOR NON-COMMERCIAL PURPOSES, SUBJECT TO PROPER CITATION OF THE ARTICLE, AUTHOR, AND PUBLISHER. EMBO J, Volume: 38, Issue: 15, First published: 27 May 2019, DOI: (10.15252/embj.2018101433)