Single-Molecule Protein Unfolding and Translocation by an ATP-Fueled Proteolytic Machine
All cells employ ATP-powered proteases for protein-quality control and regulation. In the ClpXP protease, ClpX is a AAA+ machine that recognizes specific protein substrates, unfolds these molecules, and then translocates the denatured polypeptide through a central pore and into ClpP for degradation....
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | en_US |
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Elsevier B.V.
2013
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| Online Access: | http://hdl.handle.net/1721.1/83088 https://orcid.org/0000-0002-4614-251X https://orcid.org/0000-0002-1719-5399 |
| _version_ | 1826188446742347776 |
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| author | Aubin-Tam, Marie-Eve Olivares, Adrian O. Baker, Tania Lang, Matthew J. Olivares, Adrian O. Sauer, Robert T. Sauer, Robert T |
| author2 | Massachusetts Institute of Technology. Department of Biological Engineering |
| author_facet | Massachusetts Institute of Technology. Department of Biological Engineering Aubin-Tam, Marie-Eve Olivares, Adrian O. Baker, Tania Lang, Matthew J. Olivares, Adrian O. Sauer, Robert T. Sauer, Robert T |
| author_sort | Aubin-Tam, Marie-Eve |
| collection | MIT |
| description | All cells employ ATP-powered proteases for protein-quality control and regulation. In the ClpXP protease, ClpX is a AAA+ machine that recognizes specific protein substrates, unfolds these molecules, and then translocates the denatured polypeptide through a central pore and into ClpP for degradation. Here, we use optical-trapping nanometry to probe the mechanics of enzymatic unfolding and translocation of single molecules of a multidomain substrate. Our experiments demonstrate the capacity of ClpXP and ClpX to perform mechanical work under load, reveal very fast and highly cooperative unfolding of individual substrate domains, suggest a translocation step size of 5–8 amino acids, and support a power-stroke model of denaturation in which successful enzyme-mediated unfolding of stable domains requires coincidence between mechanical pulling by the enzyme and a transient stochastic reduction in protein stability. We anticipate that single-molecule studies of the mechanical properties of other AAA+ proteolytic machines will reveal many shared features with ClpXP. |
| first_indexed | 2024-09-23T07:59:47Z |
| format | Article |
| id | mit-1721.1/83088 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T07:59:47Z |
| publishDate | 2013 |
| publisher | Elsevier B.V. |
| record_format | dspace |
| spelling | mit-1721.1/830882022-09-23T10:08:53Z Single-Molecule Protein Unfolding and Translocation by an ATP-Fueled Proteolytic Machine Aubin-Tam, Marie-Eve Olivares, Adrian O. Baker, Tania Lang, Matthew J. Olivares, Adrian O. Sauer, Robert T. Sauer, Robert T Massachusetts Institute of Technology. Department of Biological Engineering Massachusetts Institute of Technology. Department of Biology Massachusetts Institute of Technology. Department of Mechanical Engineering Aubin-Tam, Marie-Eve Olivares, Adrian O. Sauer, Robert T. Baker, Tania Lang, Matthew J. All cells employ ATP-powered proteases for protein-quality control and regulation. In the ClpXP protease, ClpX is a AAA+ machine that recognizes specific protein substrates, unfolds these molecules, and then translocates the denatured polypeptide through a central pore and into ClpP for degradation. Here, we use optical-trapping nanometry to probe the mechanics of enzymatic unfolding and translocation of single molecules of a multidomain substrate. Our experiments demonstrate the capacity of ClpXP and ClpX to perform mechanical work under load, reveal very fast and highly cooperative unfolding of individual substrate domains, suggest a translocation step size of 5–8 amino acids, and support a power-stroke model of denaturation in which successful enzyme-mediated unfolding of stable domains requires coincidence between mechanical pulling by the enzyme and a transient stochastic reduction in protein stability. We anticipate that single-molecule studies of the mechanical properties of other AAA+ proteolytic machines will reveal many shared features with ClpXP. National Science Foundation (U.S.) (Career Award 0643745) Howard Hughes Medical Institute National Institutes of Health (U.S.) (grant AI-82929) National Institutes of Health (U.S.) (grant AI-15706) 2013-12-19T20:44:06Z 2013-12-19T20:44:06Z 2011-04 Article http://purl.org/eprint/type/JournalArticle 00928674 http://hdl.handle.net/1721.1/83088 Aubin-Tam, Marie-Eve, Adrian O. Olivares, Robert T. Sauer, Tania A. Baker, and Matthew J. Lang. “Single-Molecule Protein Unfolding and Translocation by an ATP-Fueled Proteolytic Machine.” Cell 145, no. 2 (April 2011): 257-267. © 2011 Elsevier Inc. https://orcid.org/0000-0002-4614-251X https://orcid.org/0000-0002-1719-5399 en_US http://dx.doi.org/10.1016/j.cell.2011.03.036 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 B.V. Elsevier Open Archive |
| spellingShingle | Aubin-Tam, Marie-Eve Olivares, Adrian O. Baker, Tania Lang, Matthew J. Olivares, Adrian O. Sauer, Robert T. Sauer, Robert T Single-Molecule Protein Unfolding and Translocation by an ATP-Fueled Proteolytic Machine |
| title | Single-Molecule Protein Unfolding and Translocation by an ATP-Fueled Proteolytic Machine |
| title_full | Single-Molecule Protein Unfolding and Translocation by an ATP-Fueled Proteolytic Machine |
| title_fullStr | Single-Molecule Protein Unfolding and Translocation by an ATP-Fueled Proteolytic Machine |
| title_full_unstemmed | Single-Molecule Protein Unfolding and Translocation by an ATP-Fueled Proteolytic Machine |
| title_short | Single-Molecule Protein Unfolding and Translocation by an ATP-Fueled Proteolytic Machine |
| title_sort | single molecule protein unfolding and translocation by an atp fueled proteolytic machine |
| url | http://hdl.handle.net/1721.1/83088 https://orcid.org/0000-0002-4614-251X https://orcid.org/0000-0002-1719-5399 |
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