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J Mol Biol. 2017 Sep 15;429(19):2918-2930. doi: 10.1016/j.jmb.2017.08.005. Epub 2017 Aug 17.

Hsp90 Sensitivity to ADP Reveals Hidden Regulation Mechanisms.

Author information

1
Department of Biochemistry Brandeis University, Waltham, MA 02453, USA.
2
Department of Biochemistry Brandeis University, Waltham, MA 02453, USA. Electronic address: tstreet@brandeis.edu.

Abstract

The ATPase cycle of the Hsp90 molecular chaperone is essential for maintaining the stability of numerous client proteins. Extensive analysis has focused on ATP-driven conformational changes of Hsp90; however, little is known about how Hsp90 operates under physiological nucleotide conditions in which both ATP and ADP are present. By quantifying Hsp90 activity under mixed nucleotide conditions, we find dramatic differences in ADP sensitivity among Hsp90 homologs. ADP acts as a strong ATPase inhibitor of cytosol-specific Hsp90 homologs, whereas organellular Hsp90 homologs (Grp94 and TRAP1) are relatively insensitive to the presence of ADP. These results imply that an ATP/ADP heterodimer of cytosolic Hsp90 is the predominant active state under physiological nucleotide conditions. ADP inhibition of human and yeast cytosolic Hsp90 can be relieved by the cochaperone aha1. ADP inhibition of bacterial Hsp90 can be relieved by bacterial Hsp70 and an activating client protein. These results suggest that altering ADP inhibition may be a mechanism of Hsp90 regulation. To determine the molecular origin of ADP inhibition, we identify residues that preferentially stabilize either ATP or ADP. Mutations at these sites can both increase and decrease ADP inhibition. An accounting of ADP is critically important for designing and interpreting experiments with Hsp90. For example, contaminating ADP is a confounding factor in fluorescence resonance energy transfer experiments measuring arm closure rates of Hsp90. Our observations suggest that ADP at physiological levels is important to Hsp90 structure, activity, and regulation.

KEYWORDS:

ATPase; Hsp90; chaperone; inhibition; regulation

PMID:
28822683
PMCID:
PMC5610941
DOI:
10.1016/j.jmb.2017.08.005
[Indexed for MEDLINE]
Free PMC Article

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