Transmembrane helix hydrophobicity is an energetic barrier during the retrotranslocation of integral membrane ERAD substrates
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- Christopher J. Guerriero
- Department of Biological Sciences, University of Pittsburgh, Pittsburgh, PA 15260
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- Karl-Richard Reutter
- Department of Biological Sciences, University of Pittsburgh, Pittsburgh, PA 15260
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- Andrew A. Augustine
- Department of Biological Sciences, University of Pittsburgh, Pittsburgh, PA 15260
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- G. Michael Preston
- Department of Biological Sciences, University of Pittsburgh, Pittsburgh, PA 15260
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- Kurt F. Weiberth
- Department of Biological Sciences, University of Pittsburgh, Pittsburgh, PA 15260
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- Timothy D. Mackie
- Department of Biological Sciences, University of Pittsburgh, Pittsburgh, PA 15260
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- Hillary C. Cleveland-Rubeor
- Department of Biological Sciences, University of Pittsburgh, Pittsburgh, PA 15260
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- Neville P. Bethel
- Cardiovascular Research Institute, Department of Pharmaceutical Chemistry, University of California, San Francisco, San Francisco, CA 94158
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- Keith M. Callenberg
- Department of Biological Sciences, University of Pittsburgh, Pittsburgh, PA 15260
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- Kunio Nakatsukasa
- Department of Biological Sciences, University of Pittsburgh, Pittsburgh, PA 15260
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- Michael Grabe
- Cardiovascular Research Institute, Department of Pharmaceutical Chemistry, University of California, San Francisco, San Francisco, CA 94158
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- Jeffrey L. Brodsky
- Department of Biological Sciences, University of Pittsburgh, Pittsburgh, PA 15260
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- Anne Spang
- editor
説明
<jats:p>Integral membrane proteins fold inefficiently and are susceptible to turnover via the endoplasmic reticulum–associated degradation (ERAD) pathway. During ERAD, misfolded proteins are recognized by molecular chaperones, polyubiquitinated, and retrotranslocated to the cytoplasm for proteasomal degradation. Although many aspects of this pathway are defined, how transmembrane helices (TMHs) are removed from the membrane and into the cytoplasm before degradation is poorly understood. In this study, we asked whether the hydrophobic character of a TMH acts as an energetic barrier to retrotranslocation. To this end, we designed a dual-pass model ERAD substrate, Chimera A*, which contains the cytoplasmic misfolded domain from a characterized ERAD substrate, Sterile 6* (Ste6p*). We found that the degradation requirements for Chimera A* and Ste6p* are similar, but Chimera A* was retrotranslocated more efficiently than Ste6p* in an in vitro assay in which retrotranslocation can be quantified. We then constructed a series of Chimera A* variants containing synthetic TMHs with a range of ΔG values for membrane insertion. TMH hydrophobicity correlated inversely with retrotranslocation efficiency, and in all cases, retrotranslocation remained Cdc48p dependent. These findings provide insight into the energetic restrictions on the retrotranslocation reaction, as well as a new computational approach to predict retrotranslocation efficiency.</jats:p>
収録刊行物
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- Molecular Biology of the Cell
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Molecular Biology of the Cell 28 (15), 2076-2090, 2017-07-15
American Society for Cell Biology (ASCB)
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キーワード
- Adenosine Triphosphatases
- Proteasome Endopeptidase Complex
- Protein Folding
- Membranes
- Saccharomyces cerevisiae Proteins
- Ubiquitin
- Ubiquitin-Protein Ligases
- Ubiquitination
- Membrane Proteins
- Cell Cycle Proteins
- Articles
- Endoplasmic Reticulum-Associated Degradation
- Saccharomyces cerevisiae
- Endoplasmic Reticulum
- Protein Transport
- Mutation
- Protein Translocation Systems
- Hydrophobic and Hydrophilic Interactions
詳細情報 詳細情報について
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- CRID
- 1362825895530717952
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- ISSN
- 19394586
- 10591524
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- PubMed
- 28539401
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- データソース種別
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- Crossref
- OpenAIRE
