Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Experiment Videos

Structural insights into substrate binding by the molecular chaperone DnaK.

M Pellecchia1, D L Montgomery, S Y Stevens

  • 1Biophysics Research Division University of Michigan, 930 North University Avenue, Ann Arbor, Michigan 48109 USA.

Nature Structural Biology
|March 31, 2000
PubMed
Summary

The Hsp70 chaperone DnaK

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Corrigendum to "Mutations and clinical significance of calcium voltage-gated channel subunit alpha 1E (CACNA1E) in non-small cell lung cancer" [Cell Calcium 102 (2022) 102527].

Cell calcium·2024
Same author

[Clinical features and related factors of invasive pulmonary aspergillosis in patients with acute exacerbation of chronic obstructive pulmonary disease].

Zhonghua yi xue za zhi·2023
Same author

Heart Rate Response to Racquetball.

The Physician and sportsmedicine·2016
Same author

A Comparison of the Intensity of Play in Squash and Running.

The Physician and sportsmedicine·2016
Same author

BI-69A11 enhances susceptibility of colon cancer cells to mda-7/IL-24-induced growth inhibition by targeting Akt.

British journal of cancer·2014
Same author

Genetic inactivation or pharmacological inhibition of Pdk1 delays development and inhibits metastasis of Braf(V600E)::Pten(-/-) melanoma.

Oncogene·2013

Area of Science:

  • Molecular Biology
  • Structural Biology
  • Biochemistry

Background:

  • The 70 kDa heat shock protein (Hsp70) family of molecular chaperones plays critical roles in protein homeostasis.
  • Understanding how nucleotide binding regulates Hsp70 substrate affinity is a key challenge.

Purpose of the Study:

  • To investigate the mechanism by which nucleotide binding modulates substrate binding affinity in E. coli Hsp70 (DnaK).
  • To determine the structural basis for conformational changes in the DnaK substrate binding domain.

Main Methods:

  • Utilized in vivo lambda phage replication assays to assess functional activity of truncated DnaK.
  • Determined the NMR solution structure of the DnaK substrate binding domain (DnaK(393-507)) in the absence and presence of substrate.
  • Assessed conformational changes in the substrate binding domain upon substrate binding.

Related Experiment Videos

Main Results:

  • Truncated DnaK (DnaK(1-507)) retains in vivo function and in vitro communication between nucleotide and substrate binding domains.
  • The DnaK substrate binding domain (DnaK(393-507)) adopts a distinct conformation without substrate, occluding the binding site.
  • Substrate binding induces a conformational shift in the beta-domain towards previously observed structures.

Conclusions:

  • Conformational changes within the Hsp70 beta-domain are integral to the mechanism of nucleotide-modulated substrate affinity.
  • These findings provide insights into the allosteric regulation of Hsp70 molecular chaperones.