Related Experiment Video
Updated: May 29, 2026

In Situ Monitoring of Transiently Formed Molecular Chaperone Assemblies in Bacteria, Yeast, and Human Cells
Published on: September 2, 2019
Nucleotide exchange factors for Hsp70 chaperones
Heike Rampelt1, Matthias P Mayer, Bernd Bukau
1Zentrum für Molekulare Biologie, der Universität Heidelberg, DKFZ-ZMBH Alliance, Heidelberg, Germany.
This study details methods for analyzing heat shock protein 70 (Hsp70) nucleotide exchange factors (NEFs). These factors are crucial for Hsp70
Area of Science:
- Molecular Biology
- Protein Biochemistry
- Chaperone Proteins
Background:
- Heat shock protein 70 (Hsp70) chaperones regulate protein folding through their ATPase cycle.
- Nucleotide exchange factors (NEFs) modulate Hsp70 activity by accelerating ADP release, facilitating ATP rebinding and substrate release.
Purpose of the Study:
- To describe methodologies for the study and characterization of Hsp70 NEFs.
- To provide tools for understanding NEF influence on the Hsp70 ATPase cycle.
Main Methods:
- Steady-state ATPase assays to assess NEF impact on the Hsp70 ATPase cycle.
- Direct measurement of nucleotide release using labeled nucleotides for NEF identification and characterization.
Main Results:
- Steady-state ATPase assays reveal how NEFs affect Hsp70's progression through its ATPase cycle.
- Direct nucleotide release assays enable the identification and detailed characterization of NEFs.
Conclusions:
- Established methods allow for comprehensive analysis of Hsp70 NEFs.
- These techniques are vital for understanding the role of NEFs in Hsp70-mediated protein folding and cellular processes.
More Related Videos
10:24Defining Hsp33's Redox-regulated Chaperone Activity and Mapping Conformational Changes on Hsp33 Using Hydrogen-deuterium Exchange Mass Spectrometry
Published on: June 7, 2018
07:14Escherichia coli -Based Complementation Assay to Study the Chaperone Function of Heat Shock Protein 70
Published on: March 8, 2024
Related Concept Videos
Molecular Chaperones and Protein Folding
The...
Molecular Chaperones and Protein Folding
The...
Bacterial Protein Maturation
Energy to Drive Translocation
Generally, polypeptides are unfolded by two distinct...
Post-translational Translocation of Proteins to the RER
Targeting proteins to the ER
Hsp40 and Hsp70 chaperone molecules bind the translated proteins in the cytosol to prevent their folding. The chaperone binding helps to keep the signal...
Protein Complexes with Interchangeable Parts
The SCF ubiquitin ligase is a protein complex of five individual proteins. This complex attaches ubiquitin to other target proteins to mark them for degradation. In order to...