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Related Experiment Video
Updated: Sep 14, 2025

Analyzing Protein Dynamics Using Hydrogen Exchange Mass Spectrometry
Published on: November 29, 2013
ATP plays a structural role in Hsp90 function.
Michael Reidy1, Daniel C Masison2
1Laboratory of Biochemistry and Genetics, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD, USA. michael.reidy@nih.gov.
Adenosine triphosphate (ATP) binding, not hydrolysis, is essential for Hsp90 chaperone function. ATP acts as a structural linker, stabilizing the Hsp90 clamp through interactions with arginine R380.
Area of Science:
- Biochemistry
- Molecular Biology
- Structural Biology
Background:
- Heat shock protein 90 (Hsp90) is a crucial molecular chaperone.
- Hsp90 utilizes adenosine triphosphate (ATP) for its function.
- The precise role of ATP binding versus hydrolysis in Hsp90's mechanism remains unclear.
Purpose of the Study:
- To elucidate the structural role of ATP in Hsp90 chaperone function.
- To investigate the interaction between ATP and conserved residues, specifically arginine R380.
- To understand how ATP binding and hydrolysis influence Hsp90 clamp dynamics.
Main Methods:
- The study likely involved structural biology techniques (e.g., X-ray crystallography, cryo-EM) to visualize Hsp90-ATP complexes.
- Biochemical assays were probably used to assess the impact of ATP binding and hydrolysis on Hsp90 activity.
- Mutagenesis studies targeting R380 may have been employed to test its functional significance.
Main Results:
- Findings indicate that ATP binding, not hydrolysis, is critical for Hsp90's structural integrity and function.
- The gamma phosphate of ATP repositions to interact with arginine R380, stabilizing the closed chaperone clamp.
- ATP acts as a structural linker, tethering the N and M domains of Hsp90, with hydrolysis facilitating clamp reopening.
Conclusions:
- The essential role of ATP in Hsp90 function is structural, serving to stabilize the closed clamp conformation.
- Arginine R380 functions as an "arginine finger" by interacting with ATP, suggesting nucleotides can be structural elements in NTPase families.
- This work provides new insights into the mechanism of Hsp90 and the broader role of nucleotides in protein-ligand interactions.

