Dissection of Axial-Pore Loop Function during Unfolding and Translocation by a AAA+ Proteolytic Machine
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Iosefson-2015-Dissection of Axial-.pdf
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Author(s)
Iosefson, Ohad
Olivares, Adrian O.
Baker, Tania
Sauer, Robert T
Date Issued
July 2015
Journal
Cell Reports
Publisher
Elsevier
Citation
Iosefson, Ohad, Adrian O. Olivares, Tania A. Baker, and Robert T. Sauer. “Dissection of Axial-Pore Loop Function during Unfolding and Translocation by a AAA+ Proteolytic Machine.” Cell Reports 12, no. 6 (August 2015): 1032–41.
Version
Final published version
Abstract
In the axial channels of ClpX and related hexameric AAA+ protein-remodeling rings, the pore-1 loops are thought to play important roles in engaging, mechanically unfolding, and translocating protein substrates. How these loops perform these functions and whether they also prevent substrate dissociation to ensure processive degradation by AAA+ proteases are open questions. Using ClpX pore-1-loop variants, single-molecule force spectroscopy, and ensemble assays, we find that the six pore-1 loops function synchronously to grip and unfold protein substrates during a power stroke but are not important in preventing substrate slipping between power strokes. The importance of grip strength is task dependent. ClpX variants with multiple mutant pore-1 loops translocate substrates as well as the wild-type enzyme against a resisting force but show unfolding defects and a higher frequency of substrate release. These problems are magnified for more mechanically stable target proteins, supporting a threshold model of substrate gripping.
MIT Department
Massachusetts Institute of Technology. Department of Biology
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DOI of Published Version
https://doi.org/10.1016/j.celrep.2015.07.007
