XThe ClpXP protease unfolds substrates using a constant rate of pulling but different gears

Maya Sen, Rodrigo A. Maillard, Kristofor Nyquist, Piere Rodriguez-Aliaga, Steve Presse, Andreas Martin, Carlos Bustamante

Research output: Contribution to journalArticle

70 Citations (Scopus)

Abstract

ATP-dependent proteases are vital to maintain cellular protein homeostasis. Here, we study the mechanisms of force generation and intersubunit coordination in the ClpXP protease from E. coli to understand how these machines couple ATP hydrolysis to mechanical protein unfolding. Single-molecule analyses reveal that phosphate release is the force-generating step in the ATP-hydrolysis cycle and that ClpXP translocates substrate polypeptides in bursts resulting from highly coordinated conformational changes in two to four ATPase subunits. ClpXP must use its maximum successive firing capacity of four subunits to unfold stable substrates like GFP. The average dwell duration between individual bursts of translocation is constant, regardless of the number of translocating subunits, implying that ClpXP operates with constant "rpm" but uses different "gears."

Original languageEnglish (US)
JournalCell
Volume155
Issue number3
DOIs
StatePublished - Oct 24 2013
Externally publishedYes

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Gears
Hydrolysis
Peptide Hydrolases
ATP-Dependent Proteases
Adenosine Triphosphate
Protein Unfolding
Substrates
Adenosine Triphosphatases
Proteins
Homeostasis
Phosphates
Peptides
Molecules
E coli ClpXP protease

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Sen, M., Maillard, R. A., Nyquist, K., Rodriguez-Aliaga, P., Presse, S., Martin, A., & Bustamante, C. (2013). XThe ClpXP protease unfolds substrates using a constant rate of pulling but different gears. Cell, 155(3). https://doi.org/10.1016/j.cell.2013.09.022

XThe ClpXP protease unfolds substrates using a constant rate of pulling but different gears. / Sen, Maya; Maillard, Rodrigo A.; Nyquist, Kristofor; Rodriguez-Aliaga, Piere; Presse, Steve; Martin, Andreas; Bustamante, Carlos.

In: Cell, Vol. 155, No. 3, 24.10.2013.

Research output: Contribution to journalArticle

Sen, M, Maillard, RA, Nyquist, K, Rodriguez-Aliaga, P, Presse, S, Martin, A & Bustamante, C 2013, 'XThe ClpXP protease unfolds substrates using a constant rate of pulling but different gears', Cell, vol. 155, no. 3. https://doi.org/10.1016/j.cell.2013.09.022
Sen, Maya ; Maillard, Rodrigo A. ; Nyquist, Kristofor ; Rodriguez-Aliaga, Piere ; Presse, Steve ; Martin, Andreas ; Bustamante, Carlos. / XThe ClpXP protease unfolds substrates using a constant rate of pulling but different gears. In: Cell. 2013 ; Vol. 155, No. 3.
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