Effect of directional pulling on mechanical protein degradation by ATP-dependent proteolytic machines
AAA+ proteases and remodeling machines couple hydrolysis of ATP to mechanical unfolding and translocation of proteins following recognition of sequence tags called degrons. Here, we use single-molecule optical trapping to determine the mechanochemistry of two AAA+ proteases, Escherichia coli ClpXP a...
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| Format: | Article |
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National Academy of Sciences (U.S.)
2018
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| Online Access: | http://hdl.handle.net/1721.1/114914 https://orcid.org/0000-0001-9751-9535 https://orcid.org/0000-0002-2246-2674 https://orcid.org/0000-0002-1719-5399 |
| _version_ | 1826190051137028096 |
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| author | Olivares, Adrian O. Kotamarthi, Hema Chandra Stein, Benjamin Joseph Sauer, Robert T. Baker, Tania |
| author2 | Massachusetts Institute of Technology. Department of Biology |
| author_facet | Massachusetts Institute of Technology. Department of Biology Olivares, Adrian O. Kotamarthi, Hema Chandra Stein, Benjamin Joseph Sauer, Robert T. Baker, Tania |
| author_sort | Olivares, Adrian O. |
| collection | MIT |
| description | AAA+ proteases and remodeling machines couple hydrolysis of ATP to mechanical unfolding and translocation of proteins following recognition of sequence tags called degrons. Here, we use single-molecule optical trapping to determine the mechanochemistry of two AAA+ proteases, Escherichia coli ClpXP and ClpAP, as they unfold and translocate substrates containing multiple copies of the titin[superscript I27] domain during degradation initiated from the N terminus. Previous studies characterized degradation of related substrates with C-terminal degrons. We find that ClpXP and ClpAP unfold the wild-type titin I27 domain and a destabilized variant far more rapidly when pulling from the N terminus, whereas translocation speed is reduced only modestly in the N-to-C direction. These measurements establish the role of directionality in mechanical protein degradation, show that degron placement can change whether unfolding or translocation is rate limiting, and establish that one or a few power strokes are sufficient to unfold some protein domains. Keywords:protein degradation; AAA+ proteases; directional unfolding; AAA+ motors |
| first_indexed | 2024-09-23T08:34:17Z |
| format | Article |
| id | mit-1721.1/114914 |
| institution | Massachusetts Institute of Technology |
| last_indexed | 2024-09-23T08:34:17Z |
| publishDate | 2018 |
| publisher | National Academy of Sciences (U.S.) |
| record_format | dspace |
| spelling | mit-1721.1/1149142022-09-30T09:41:18Z Effect of directional pulling on mechanical protein degradation by ATP-dependent proteolytic machines Olivares, Adrian O. Kotamarthi, Hema Chandra Stein, Benjamin Joseph Sauer, Robert T. Baker, Tania Massachusetts Institute of Technology. Department of Biology Olivares, Adrian O. Kotamarthi, Hema Chandra Stein, Benjamin Joseph Sauer, Robert T. Baker, Tania AAA+ proteases and remodeling machines couple hydrolysis of ATP to mechanical unfolding and translocation of proteins following recognition of sequence tags called degrons. Here, we use single-molecule optical trapping to determine the mechanochemistry of two AAA+ proteases, Escherichia coli ClpXP and ClpAP, as they unfold and translocate substrates containing multiple copies of the titin[superscript I27] domain during degradation initiated from the N terminus. Previous studies characterized degradation of related substrates with C-terminal degrons. We find that ClpXP and ClpAP unfold the wild-type titin I27 domain and a destabilized variant far more rapidly when pulling from the N terminus, whereas translocation speed is reduced only modestly in the N-to-C direction. These measurements establish the role of directionality in mechanical protein degradation, show that degron placement can change whether unfolding or translocation is rate limiting, and establish that one or a few power strokes are sufficient to unfold some protein domains. Keywords:protein degradation; AAA+ proteases; directional unfolding; AAA+ motors National Institutes of Health (U.S.) (Grant GM-101988) National Institutes of Health (U.S.) (Grant AI-15706) 2018-04-24T13:35:36Z 2018-04-24T13:35:36Z 2017-07 2017-05 2018-04-20T13:11:57Z Article http://purl.org/eprint/type/ConferencePaper 0027-8424 1091-6490 http://hdl.handle.net/1721.1/114914 Olivares, Adrian O. et al. “Effect of Directional Pulling on Mechanical Protein Degradation by ATP-Dependent Proteolytic Machines.” Proceedings of the National Academy of Sciences 114, 31 (July 2017): E6306–E6313 © 2017 National Academy of Sciences https://orcid.org/0000-0001-9751-9535 https://orcid.org/0000-0002-2246-2674 https://orcid.org/0000-0002-1719-5399 http://dx.doi.org/10.1073/PNAS.1707794114 Proceedings of the National Academy of Sciences 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 National Academy of Sciences (U.S.) National Academy of Sciences |
| spellingShingle | Olivares, Adrian O. Kotamarthi, Hema Chandra Stein, Benjamin Joseph Sauer, Robert T. Baker, Tania Effect of directional pulling on mechanical protein degradation by ATP-dependent proteolytic machines |
| title | Effect of directional pulling on mechanical protein degradation by ATP-dependent proteolytic machines |
| title_full | Effect of directional pulling on mechanical protein degradation by ATP-dependent proteolytic machines |
| title_fullStr | Effect of directional pulling on mechanical protein degradation by ATP-dependent proteolytic machines |
| title_full_unstemmed | Effect of directional pulling on mechanical protein degradation by ATP-dependent proteolytic machines |
| title_short | Effect of directional pulling on mechanical protein degradation by ATP-dependent proteolytic machines |
| title_sort | effect of directional pulling on mechanical protein degradation by atp dependent proteolytic machines |
| url | http://hdl.handle.net/1721.1/114914 https://orcid.org/0000-0001-9751-9535 https://orcid.org/0000-0002-2246-2674 https://orcid.org/0000-0002-1719-5399 |
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