Symmetry broken and rebroken during the ATP hydrolysis cycle of the mitochondrial Hsp90 TRAP1
Hsp90 is a homodimeric ATP-dependent molecular chaperone that remodels its substrate ‘client’ proteins, facilitating their folding and activating them for biological function. Despite decades of research, the mechanism connecting ATP hydrolysis and chaperone function remains elusive. Particularly pu...
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eLife Sciences Publications Ltd
2017-07-01
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Series: | eLife |
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Online Access: | https://elifesciences.org/articles/25235 |
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author | Daniel Elnatan Miguel Betegon Yanxin Liu Theresa Ramelot Michael A Kennedy David A Agard |
author_facet | Daniel Elnatan Miguel Betegon Yanxin Liu Theresa Ramelot Michael A Kennedy David A Agard |
author_sort | Daniel Elnatan |
collection | DOAJ |
description | Hsp90 is a homodimeric ATP-dependent molecular chaperone that remodels its substrate ‘client’ proteins, facilitating their folding and activating them for biological function. Despite decades of research, the mechanism connecting ATP hydrolysis and chaperone function remains elusive. Particularly puzzling has been the apparent lack of cooperativity in hydrolysis of the ATP in each protomer. A crystal structure of the mitochondrial Hsp90, TRAP1, revealed that the catalytically active state is closed in a highly strained asymmetric conformation. This asymmetry, unobserved in other Hsp90 homologs, is due to buckling of one of the protomers and is most pronounced at the broadly conserved client-binding region. Here, we show that rather than being cooperative or independent, ATP hydrolysis on the two protomers is sequential and deterministic. Moreover, dimer asymmetry sets up differential hydrolysis rates for each protomer, such that the buckled conformation favors ATP hydrolysis. Remarkably, after the first hydrolysis, the dimer undergoes a flip in the asymmetry while remaining in a closed state for the second hydrolysis. From these results, we propose a model where direct coupling of ATP hydrolysis and conformational flipping rearranges client-binding sites, providing a paradigm of how energy from ATP hydrolysis can be used for client remodeling. |
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institution | Directory Open Access Journal |
issn | 2050-084X |
language | English |
last_indexed | 2024-12-10T04:07:22Z |
publishDate | 2017-07-01 |
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spelling | doaj.art-dec2959eea6d4b2abe575a0f8f110a032022-12-22T02:02:49ZengeLife Sciences Publications LtdeLife2050-084X2017-07-01610.7554/eLife.25235Symmetry broken and rebroken during the ATP hydrolysis cycle of the mitochondrial Hsp90 TRAP1Daniel Elnatan0https://orcid.org/0000-0002-8359-0522Miguel Betegon1https://orcid.org/0000-0001-7625-6190Yanxin Liu2https://orcid.org/0000-0002-2253-3698Theresa Ramelot3Michael A Kennedy4David A Agard5https://orcid.org/0000-0003-3512-695XDepartment of Biochemistry and Biophysics, Howard Hughes Medical Institute, University of California, San Francisco, United States; Tetrad Graduate program, University of California, San Francisco, United StatesDepartment of Biochemistry and Biophysics, Howard Hughes Medical Institute, University of California, San Francisco, United States; Biophysics Graduate program, University of California, San Francisco, United StatesDepartment of Biochemistry and Biophysics, Howard Hughes Medical Institute, University of California, San Francisco, United StatesDepartment of Chemistry and Biochemistry, Miami University, Oxford, United StatesDepartment of Chemistry and Biochemistry, Miami University, Oxford, United StatesDepartment of Biochemistry and Biophysics, Howard Hughes Medical Institute, University of California, San Francisco, United StatesHsp90 is a homodimeric ATP-dependent molecular chaperone that remodels its substrate ‘client’ proteins, facilitating their folding and activating them for biological function. Despite decades of research, the mechanism connecting ATP hydrolysis and chaperone function remains elusive. Particularly puzzling has been the apparent lack of cooperativity in hydrolysis of the ATP in each protomer. A crystal structure of the mitochondrial Hsp90, TRAP1, revealed that the catalytically active state is closed in a highly strained asymmetric conformation. This asymmetry, unobserved in other Hsp90 homologs, is due to buckling of one of the protomers and is most pronounced at the broadly conserved client-binding region. Here, we show that rather than being cooperative or independent, ATP hydrolysis on the two protomers is sequential and deterministic. Moreover, dimer asymmetry sets up differential hydrolysis rates for each protomer, such that the buckled conformation favors ATP hydrolysis. Remarkably, after the first hydrolysis, the dimer undergoes a flip in the asymmetry while remaining in a closed state for the second hydrolysis. From these results, we propose a model where direct coupling of ATP hydrolysis and conformational flipping rearranges client-binding sites, providing a paradigm of how energy from ATP hydrolysis can be used for client remodeling.https://elifesciences.org/articles/25235Hsp90TRAP1chaperoneATP hydrolysishomodimerasymmetry |
spellingShingle | Daniel Elnatan Miguel Betegon Yanxin Liu Theresa Ramelot Michael A Kennedy David A Agard Symmetry broken and rebroken during the ATP hydrolysis cycle of the mitochondrial Hsp90 TRAP1 eLife Hsp90 TRAP1 chaperone ATP hydrolysis homodimer asymmetry |
title | Symmetry broken and rebroken during the ATP hydrolysis cycle of the mitochondrial Hsp90 TRAP1 |
title_full | Symmetry broken and rebroken during the ATP hydrolysis cycle of the mitochondrial Hsp90 TRAP1 |
title_fullStr | Symmetry broken and rebroken during the ATP hydrolysis cycle of the mitochondrial Hsp90 TRAP1 |
title_full_unstemmed | Symmetry broken and rebroken during the ATP hydrolysis cycle of the mitochondrial Hsp90 TRAP1 |
title_short | Symmetry broken and rebroken during the ATP hydrolysis cycle of the mitochondrial Hsp90 TRAP1 |
title_sort | symmetry broken and rebroken during the atp hydrolysis cycle of the mitochondrial hsp90 trap1 |
topic | Hsp90 TRAP1 chaperone ATP hydrolysis homodimer asymmetry |
url | https://elifesciences.org/articles/25235 |
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