Substrate-translocating loops regulate mechanochemical coupling and power production in AAA+ protease ClpXP.
ATP-dependent proteases of the AAA+ family, including Escherichia coli ClpXP and the eukaryotic proteasome, contribute to maintenance of cellular proteostasis. ClpXP unfolds and translocates substrates into an internal degradation chamber, using cycles of alternating dwell and burst phases. The ClpX motor performs chemical transformations during the dwell and translocates the substrate in increments of 1-4 nm during the burst, but the processes occurring during these phases remain unknown. Here we characterized the complete mechanochemical cycle of ClpXP, showing that ADP release and ATP binding occur nonsequentially during the dwell, whereas ATP hydrolysis and phosphate release occur during the burst. The highly conserved translocating loops within the ClpX pore are optimized to maximize motor power generation, the coupling between chemical and mechanical tasks, and the efficiency of protein processing. Conformational resetting of these loops between consecutive bursts appears to determine ADP release from individual ATPase subunits and the overall duration of the motor's cycle.
Jason L. Choy Laboratory of Single-Molecule Biophysics, University of California at Berkeley, Berkeley, California, USA.|Biophysics Graduate Group, University of California at Berkeley, Berkeley, California, USA.|Institute for Quantitative Biosciences-QB3, University of California at Berkeley, Berkeley, California, USA.
Piere
P
Rodriguez-Aliaga
Jason L. Choy Laboratory of Single-Molecule Biophysics, University of California at Berkeley, Berkeley, California, USA.|Institute for Quantitative Biosciences-QB3, University of California at Berkeley, Berkeley, California, USA.
Luis
L
Ramirez
Jason L. Choy Laboratory of Single-Molecule Biophysics, University of California at Berkeley, Berkeley, California, USA.
Frank
F
Kim
Jason L. Choy Laboratory of Single-Molecule Biophysics, University of California at Berkeley, Berkeley, California, USA.|Biophysics Graduate Group, University of California at Berkeley, Berkeley, California, USA.|Institute for Quantitative Biosciences-QB3, University of California at Berkeley, Berkeley, California, USA.|Department of Molecular &Cell Biology, University of California at Berkeley, Berkeley, California, USA.|Department of Chemistry, University of California at Berkeley, Berkeley, California, USA.|Department of Physics, University of California at Berkeley, Berkeley, California, USA.|Howard Hughes Medical Institute, University of California at Berkeley, Berkeley, California, USA.|Kavli Energy Nanosciences Institute, University of California at Berkeley, Berkeley, California, USA.
Carlos
C
Bustamante
Biophysics Graduate Group, University of California at Berkeley, Berkeley, California, USA.|Institute for Quantitative Biosciences-QB3, University of California at Berkeley, Berkeley, California, USA.|Department of Molecular &Cell Biology, University of California at Berkeley, Berkeley, California, USA.|Howard Hughes Medical Institute, University of California at Berkeley, Berkeley, California, USA.
Andreas
A
Martin
D000074183:ATPases Associated with Diverse Cellular Activities; D000244:Adenosine Diphosphate / Q000378:metabolism; D000251:Adenosine Triphosphatases / Q000737:chemistry / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000378:metabolism*; D049071:Endopeptidase Clp / Q000737:chemistry / Q000378:metabolism*; D004926:Escherichia coli / Q000737:chemistry / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000737:chemistry / Q000378:metabolism*; D006868:Hydrolysis; D007700:Kinetics; D008958:Models, Molecular; D018832:Molecular Chaperones / Q000737:chemistry / Q000378:metabolism; D011485:Protein Binding; D011487:Protein Conformation; D058767:Protein Unfolding; D013379:Substrate Specificity
D029968:Escherichia coli Proteins; D018832:Molecular Chaperones; D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate; C483573:ClpXP protease, E coli; D049071:Endopeptidase Clp; D000251:Adenosine Triphosphatases; C088072:ClpX protein, E coli; D000074183:ATPases Associated with Diverse Cellular Activities
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