Stochastic but Highly Coordinated Protein Unfolding and Translocation by the ClpXP Proteolytic Machine
ClpXP and other AAA+ proteases recognize, mechanically unfold, and translocate target proteins into a chamber for proteolysis. It is not known whether these remarkable molecular machines operate by a stochastic or sequential mechanism or how power strokes relate to the ATP-hydrolysis cycle. Single-m...
| Main Authors: | , , , , , , , , , |
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| Format: | Article |
| Language: | en_US |
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Elsevier
2017
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| Online Access: | http://hdl.handle.net/1721.1/106303 https://orcid.org/0000-0002-1081-285X https://orcid.org/0000-0002-9309-8662 https://orcid.org/0000-0002-1719-5399 |
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| author | Cordova, Juan Carlos Aubin-Tam, Marie-Eve Lang, Matthew J. Olivares, Adrian O. Shin, Yongdae Calmat, Stephane Geraldine Michele Schmitz, Karl Robert Baker, Tania Sauer, Robert T. Stinson, Benjamin Michael |
| author2 | Massachusetts Institute of Technology. Department of Biology |
| author_facet | Massachusetts Institute of Technology. Department of Biology Cordova, Juan Carlos Aubin-Tam, Marie-Eve Lang, Matthew J. Olivares, Adrian O. Shin, Yongdae Calmat, Stephane Geraldine Michele Schmitz, Karl Robert Baker, Tania Sauer, Robert T. Stinson, Benjamin Michael |
| author_sort | Cordova, Juan Carlos |
| collection | MIT |
| description | ClpXP and other AAA+ proteases recognize, mechanically unfold, and translocate target proteins into a chamber for proteolysis. It is not known whether these remarkable molecular machines operate by a stochastic or sequential mechanism or how power strokes relate to the ATP-hydrolysis cycle. Single-molecule optical trapping allows ClpXP unfolding to be directly visualized and reveals translocation steps of ∼1–4 nm in length, but how these activities relate to solution degradation and the physical properties of substrate proteins remains unclear. By studying single-molecule degradation using different multidomain substrates and ClpXP variants, we answer many of these questions and provide evidence for stochastic unfolding and translocation. We also present a mechanochemical model that accounts for single-molecule, biochemical, and structural results for our observation of enzymatic memory in translocation stepping, for the kinetics of translocation steps of different sizes, and for probabilistic but highly coordinated subunit activity within the ClpX ring. |
| first_indexed | 2024-09-23T08:01:47Z |
| format | Article |
| id | mit-1721.1/106303 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T08:01:47Z |
| publishDate | 2017 |
| publisher | Elsevier |
| record_format | dspace |
| spelling | mit-1721.1/1063032022-09-23T10:25:40Z Stochastic but Highly Coordinated Protein Unfolding and Translocation by the ClpXP Proteolytic Machine Cordova, Juan Carlos Aubin-Tam, Marie-Eve Lang, Matthew J. Olivares, Adrian O. Shin, Yongdae Calmat, Stephane Geraldine Michele Schmitz, Karl Robert Baker, Tania Sauer, Robert T. Stinson, Benjamin Michael Massachusetts Institute of Technology. Department of Biology Massachusetts Institute of Technology. Department of Mechanical Engineering Olivares, Adrian O. Shin, Yongdae Calmat, Stephane Geraldine Michele Schmitz, Karl Robert Baker, Tania Sauer, Robert T. Stinson, Benjamin Michael ClpXP and other AAA+ proteases recognize, mechanically unfold, and translocate target proteins into a chamber for proteolysis. It is not known whether these remarkable molecular machines operate by a stochastic or sequential mechanism or how power strokes relate to the ATP-hydrolysis cycle. Single-molecule optical trapping allows ClpXP unfolding to be directly visualized and reveals translocation steps of ∼1–4 nm in length, but how these activities relate to solution degradation and the physical properties of substrate proteins remains unclear. By studying single-molecule degradation using different multidomain substrates and ClpXP variants, we answer many of these questions and provide evidence for stochastic unfolding and translocation. We also present a mechanochemical model that accounts for single-molecule, biochemical, and structural results for our observation of enzymatic memory in translocation stepping, for the kinetics of translocation steps of different sizes, and for probabilistic but highly coordinated subunit activity within the ClpX ring. National Science Foundation (U.S.) ((Career Award 0643745 and MCB-1330792) National Institutes of Health (U.S.) (Grant GM-101988) Singapore-MIT Alliance for Research and Technology (SMART) Howard Hughes Medical Institute Samsung Foundation of Culture (Samsung Scholarship) Massachusetts Institute of Technology. Poitras Pre-Doctoral Fellowship 2017-01-09T20:13:22Z 2017-01-09T20:13:22Z 2014-07 2014-04 Article http://purl.org/eprint/type/JournalArticle 0092-8674 1097-4172 http://hdl.handle.net/1721.1/106303 Cordova, Juan Carlos et al. “Stochastic but Highly Coordinated Protein Unfolding and Translocation by the ClpXP Proteolytic Machine.” Cell 158.3 (2014): 647–658. https://orcid.org/0000-0002-1081-285X https://orcid.org/0000-0002-9309-8662 https://orcid.org/0000-0002-1719-5399 en_US http://dx.doi.org/10.1016/j.cell.2014.05.043 Cell Creative Commons Attribution-NonCommercial-NoDerivs License http://creativecommons.org/licenses/by-nc-nd/4.0/ application/pdf Elsevier PMC |
| spellingShingle | Cordova, Juan Carlos Aubin-Tam, Marie-Eve Lang, Matthew J. Olivares, Adrian O. Shin, Yongdae Calmat, Stephane Geraldine Michele Schmitz, Karl Robert Baker, Tania Sauer, Robert T. Stinson, Benjamin Michael Stochastic but Highly Coordinated Protein Unfolding and Translocation by the ClpXP Proteolytic Machine |
| title | Stochastic but Highly Coordinated Protein Unfolding and Translocation by the ClpXP Proteolytic Machine |
| title_full | Stochastic but Highly Coordinated Protein Unfolding and Translocation by the ClpXP Proteolytic Machine |
| title_fullStr | Stochastic but Highly Coordinated Protein Unfolding and Translocation by the ClpXP Proteolytic Machine |
| title_full_unstemmed | Stochastic but Highly Coordinated Protein Unfolding and Translocation by the ClpXP Proteolytic Machine |
| title_short | Stochastic but Highly Coordinated Protein Unfolding and Translocation by the ClpXP Proteolytic Machine |
| title_sort | stochastic but highly coordinated protein unfolding and translocation by the clpxp proteolytic machine |
| url | http://hdl.handle.net/1721.1/106303 https://orcid.org/0000-0002-1081-285X https://orcid.org/0000-0002-9309-8662 https://orcid.org/0000-0002-1719-5399 |
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