The conserved arginine 380 of Hsp90 is not a catalytic residue, but stabilizes the closed conformation required for ATP hydrolysis
dc.contributor.author | Cunningham, Christian N. | en_US |
dc.contributor.author | Southworth, Daniel R. | en_US |
dc.contributor.author | Krukenberg, Kristin A. | en_US |
dc.contributor.author | Agard, David A. | en_US |
dc.date.accessioned | 2012-08-09T14:56:00Z | |
dc.date.available | 2013-10-01T17:06:31Z | en_US |
dc.date.issued | 2012-08 | en_US |
dc.identifier.citation | Cunningham, Christian N.; Southworth, Daniel R.; Krukenberg, Kristin A.; Agard, David A. (2012). "The conserved arginine 380 of Hsp90 is not a catalytic residue, but stabilizes the closed conformation required for ATP hydrolysis." Protein Science 21(8): 1162-1171. <http://hdl.handle.net/2027.42/92411> | en_US |
dc.identifier.issn | 0961-8368 | en_US |
dc.identifier.issn | 1469-896X | en_US |
dc.identifier.uri | https://hdl.handle.net/2027.42/92411 | |
dc.description.abstract | Hsp90, a dimeric ATP‐dependent molecular chaperone, is required for the folding and activation of numerous essential substrate “client” proteins including nuclear receptors, cell cycle kinases, and telomerase. Fundamental to its mechanism is an ensemble of dramatically different conformational states that result from nucleotide binding and hydrolysis and distinct sets of interdomain interactions. Previous structural and biochemical work identified a conserved arginine residue (R380 in yeast) in the Hsp90 middle domain (MD) that is required for wild type hydrolysis activity in yeast, and hence proposed to be a catalytic residue. As part of our investigations on the origins of species‐specific differences in Hsp90 conformational dynamics we probed the role of this MD arginine in bacterial, yeast, and human Hsp90s using a combination of structural and functional approaches. While the R380A mutation compromised ATPase activity in all three homologs, the impact on ATPase activity was both variable and much more modest (2–7 fold) than the mutation of an active site glutamate (40 fold) known to be required for hydrolysis. Single particle electron microscopy and small‐angle X‐ray scattering revealed that, for all Hsp90s, mutation of this arginine abrogated the ability to form the closed “ATP” conformational state in response to AMPPNP binding. Taken together with previous mutagenesis data exploring intra‐ and intermonomer interactions, these new data suggest that R380 does not directly participate in the hydrolysis reaction as a catalytic residue, but instead acts as an ATP‐sensor to stabilize an NTD‐MD conformation required for efficient ATP hydrolysis. | en_US |
dc.publisher | Wiley Subscription Services, Inc., A Wiley Company | en_US |
dc.subject.other | Hsp90 | en_US |
dc.subject.other | Conformational Dynamics | en_US |
dc.subject.other | Catalysis | en_US |
dc.subject.other | Interactions | en_US |
dc.subject.other | ATP Hydrolysis | en_US |
dc.title | The conserved arginine 380 of Hsp90 is not a catalytic residue, but stabilizes the closed conformation required for ATP hydrolysis | en_US |
dc.type | Article | en_US |
dc.rights.robots | IndexNoFollow | en_US |
dc.subject.hlbsecondlevel | Biological Chemistry | en_US |
dc.subject.hlbtoplevel | Health Sciences | en_US |
dc.description.peerreviewed | Peer Reviewed | en_US |
dc.contributor.affiliationum | Department of Biological Chemistry, Life Sciences Institute, University of Michigan, 210 Washtenaw Ave, Ann Arbor, MI 48109 | en_US |
dc.contributor.affiliationother | Department of Systems Biology, Harvard Medical School, 200 Longwood Ave, Warren Alpert 536, Boston, MA 02115 | en_US |
dc.contributor.affiliationother | Graduate Group in Biophysics, University of California, San Francisco, California 94158 | en_US |
dc.contributor.affiliationother | Department of Biochemistry and Biophysics, Howard Hughes Medical Institute, University of California, San Francisco, California 94158 | en_US |
dc.contributor.affiliationother | Graduate Program in Chemistry and Chemical Biology, University of California, San Francisco, California 94158 | en_US |
dc.contributor.affiliationother | Early Discovery Biochemistry, Genentech, Inc., 1 DNA Way, MS27, South San Francisco, CA 94080 | en_US |
dc.contributor.affiliationother | 600 16th St., MC2240 Room S412D, San Francisco, CA 94158 | en_US |
dc.identifier.pmid | 22653663 | en_US |
dc.description.bitstreamurl | http://deepblue.lib.umich.edu/bitstream/2027.42/92411/1/2103_ftp.pdf | |
dc.identifier.doi | 10.1002/pro.2103 | en_US |
dc.identifier.source | Protein Science | en_US |
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dc.owningcollname | Interdisciplinary and Peer-Reviewed |
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