Covalent Linkage of Distinct Substrate Degrons Controls Assembly and Disassembly of DegP Proteolytic Cages
Protein quality control requires careful regulation of intracellular proteolysis. For DegP, a periplasmic protease, substrates promote assembly of inactive hexamers into proteolytically active cages with 12, 18, 24, or 30 subunits. Here, we show that sensitive activation and cage assembly require co...
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| Format: | Article |
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Elsevier
2014
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| Online Access: | http://hdl.handle.net/1721.1/92350 https://orcid.org/0000-0002-1719-5399 |
| _version_ | 1826214240432685056 |
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| author | Kim, Seokhee Grant, Robert A Sauer, Robert T |
| author2 | Massachusetts Institute of Technology. Department of Biology |
| author_facet | Massachusetts Institute of Technology. Department of Biology Kim, Seokhee Grant, Robert A Sauer, Robert T |
| author_sort | Kim, Seokhee |
| collection | MIT |
| description | Protein quality control requires careful regulation of intracellular proteolysis. For DegP, a periplasmic protease, substrates promote assembly of inactive hexamers into proteolytically active cages with 12, 18, 24, or 30 subunits. Here, we show that sensitive activation and cage assembly require covalent linkage of distinct substrate sequences that affect degradation (degrons). One degron binds the DegP active site, and another degron binds a separate tethering site in PDZ1 in the crystal structure of a substrate-bound DegP dodecamer. FRET experiments demonstrate that active cages assemble rapidly in a reaction that is positively cooperative in substrate concentration, remain stably assembled while uncleaved substrate is present, and dissociate once degradation is complete. Thus, the energy of binding of linked substrate degrons drives assembly of the proteolytic machine responsible for subsequent degradation. Substrate cleavage and depletion results in disassembly, ensuring that DegP is proteolytically active only when sufficient quantities of protein substrates are present. |
| first_indexed | 2024-09-23T16:02:21Z |
| format | Article |
| id | mit-1721.1/92350 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T16:02:21Z |
| publishDate | 2014 |
| publisher | Elsevier |
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| spelling | mit-1721.1/923502022-10-02T05:52:17Z Covalent Linkage of Distinct Substrate Degrons Controls Assembly and Disassembly of DegP Proteolytic Cages Kim, Seokhee Grant, Robert A Sauer, Robert T Massachusetts Institute of Technology. Department of Biology Grant, Robert A. Sauer, Robert T. Kim, Seokhee Protein quality control requires careful regulation of intracellular proteolysis. For DegP, a periplasmic protease, substrates promote assembly of inactive hexamers into proteolytically active cages with 12, 18, 24, or 30 subunits. Here, we show that sensitive activation and cage assembly require covalent linkage of distinct substrate sequences that affect degradation (degrons). One degron binds the DegP active site, and another degron binds a separate tethering site in PDZ1 in the crystal structure of a substrate-bound DegP dodecamer. FRET experiments demonstrate that active cages assemble rapidly in a reaction that is positively cooperative in substrate concentration, remain stably assembled while uncleaved substrate is present, and dissociate once degradation is complete. Thus, the energy of binding of linked substrate degrons drives assembly of the proteolytic machine responsible for subsequent degradation. Substrate cleavage and depletion results in disassembly, ensuring that DegP is proteolytically active only when sufficient quantities of protein substrates are present. National Institutes of Health (U.S.) (Grant AI-16892) United States. Dept. of Energy. Office of Basic Energy Sciences (Contract DE-AC02-06CH11357) National Center for Research Resources (U.S.) (Award RR-15301) 2014-12-16T21:18:54Z 2014-12-16T21:18:54Z 2011-03 2010-12 Article http://purl.org/eprint/type/JournalArticle 00928674 1097-4172 http://hdl.handle.net/1721.1/92350 Kim, Seokhee, Robert A. Grant, and Robert T. Sauer. “Covalent Linkage of Distinct Substrate Degrons Controls Assembly and Disassembly of DegP Proteolytic Cages.” Cell 145, no. 1 (April 2011): 67–78. © 2011 Elsevier Inc. https://orcid.org/0000-0002-1719-5399 en_US http://dx.doi.org/10.1016/j.cell.2011.02.024 Cell Article is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use. application/pdf Elsevier Elsevier |
| spellingShingle | Kim, Seokhee Grant, Robert A Sauer, Robert T Covalent Linkage of Distinct Substrate Degrons Controls Assembly and Disassembly of DegP Proteolytic Cages |
| title | Covalent Linkage of Distinct Substrate Degrons Controls Assembly and Disassembly of DegP Proteolytic Cages |
| title_full | Covalent Linkage of Distinct Substrate Degrons Controls Assembly and Disassembly of DegP Proteolytic Cages |
| title_fullStr | Covalent Linkage of Distinct Substrate Degrons Controls Assembly and Disassembly of DegP Proteolytic Cages |
| title_full_unstemmed | Covalent Linkage of Distinct Substrate Degrons Controls Assembly and Disassembly of DegP Proteolytic Cages |
| title_short | Covalent Linkage of Distinct Substrate Degrons Controls Assembly and Disassembly of DegP Proteolytic Cages |
| title_sort | covalent linkage of distinct substrate degrons controls assembly and disassembly of degp proteolytic cages |
| url | http://hdl.handle.net/1721.1/92350 https://orcid.org/0000-0002-1719-5399 |
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