Escherichia coli caseinolytic protease (Clp)B is a hexameric AAA+ [expanded superfamily of AAA (ATPase associated with various cellular activities)] enzyme that has the unique ability to catalyse protein disaggregation. Such enzymes are essential for proteome maintenance. Based on structural comparisons to homologous enzymes involved in ATP-dependent proteolysis and clever protein engineering strategies, it has been reported that ClpB translocates polypeptide through its axial channel. Using single-turnover fluorescence and anisotropy experiments we show that ClpB is a non-processive polypeptide translocase that catalyses disaggregation by taking one or two translocation steps followed by rapid dissociation. Using single-turnover FRET experiments we show that ClpB containing the IGL loop from ClpA does not translocate substrate through its axial channel and into ClpP for proteolytic degradation. Rather, ClpB containing the IGL loop dysregulates ClpP leading to non-specific proteolysis reminiscent of ADEP (acyldepsipeptide) dysregulation. Our results support a molecular mechanism where ClpB catalyses protein disaggregation by tugging and releasing exposed tails or loops.
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Research Article|
August 06 2015
Escherichia coli ClpB is a non-processive polypeptide translocase
Tao Li;
Tao Li
*Department of Chemistry, The University of Alabama at Birmingham (UAB), Birmingham AL, U.S.A.
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Clarissa L. Weaver;
Clarissa L. Weaver
*Department of Chemistry, The University of Alabama at Birmingham (UAB), Birmingham AL, U.S.A.
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Jiabei Lin;
Jiabei Lin
*Department of Chemistry, The University of Alabama at Birmingham (UAB), Birmingham AL, U.S.A.
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Elizabeth C. Duran;
Elizabeth C. Duran
*Department of Chemistry, The University of Alabama at Birmingham (UAB), Birmingham AL, U.S.A.
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Justin M. Miller;
Justin M. Miller
*Department of Chemistry, The University of Alabama at Birmingham (UAB), Birmingham AL, U.S.A.
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Aaron L. Lucius
Aaron L. Lucius
1
*Department of Chemistry, The University of Alabama at Birmingham (UAB), Birmingham AL, U.S.A.
1To whom correspondence should be addressed (email allucius@uab.edu).
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Publisher: Portland Press Ltd
Received:
December 02 2014
Revision Received:
June 04 2015
Accepted:
June 05 2015
Accepted Manuscript online:
June 11 2015
Online ISSN: 1470-8728
Print ISSN: 0264-6021
© 2015 Authors; published by Portland Press Limited
2015
Biochem J (2015) 470 (1): 39–52.
Article history
Received:
December 02 2014
Revision Received:
June 04 2015
Accepted:
June 05 2015
Accepted Manuscript online:
June 11 2015
Citation
Tao Li, Clarissa L. Weaver, Jiabei Lin, Elizabeth C. Duran, Justin M. Miller, Aaron L. Lucius; Escherichia coli ClpB is a non-processive polypeptide translocase. Biochem J 15 August 2015; 470 (1): 39–52. doi: https://doi.org/10.1042/BJ20141457
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