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

Sequence-dependent peptide binding orientation by the molecular chaperone DnaK.

Tim L Tapley1, Jill R Cupp-Vickery, Larry E Vickery

  • 1Department of Physiology and Biophysics, University of California, Irvine, California 92697, USA.

Biochemistry
|September 15, 2005
PubMed
Summary

Molecular chaperones like Hsp70 (Heat Shock Protein 70) can bind peptides in either direction. Central peptide residues, not charged flanking regions, determine this binding orientation in E. coli 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

Pharmacology and Mechanism of Action of Suzetrigine, a Potent and Selective Na<sub>V</sub>1.8 Pain Signal Inhibitor for the Treatment of Moderate to Severe Pain.

Pain and therapy·2025
Same author

Facilitated transfer of IscU-[2Fe2S] clusters by chaperone-mediated ligand exchange.

Biochemistry·2011
Same author

Three hydrophobic amino acids in Escherichia coli HscB make the greatest contribution to the stability of the HscB-IscU complex.

BMC biochemistry·2011
Same author

Structure and dynamics of the iron-sulfur cluster assembly scaffold protein IscU and its interaction with the cochaperone HscB.

Biochemistry·2009
Same author

Studies on the mechanism of catalysis of iron-sulfur cluster transfer from IscU[2Fe2S] by HscA/HscB chaperones.

Biochemistry·2008
Same author

Solution structure of the iron-sulfur cluster cochaperone HscB and its binding surface for the iron-sulfur assembly scaffold protein IscU.

Biochemistry·2008

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Structural Biology

Background:

  • Hsp70 molecular chaperones are essential for protein homeostasis.
  • Their interaction with diverse polypeptide substrates is crucial, but binding orientation is poorly understood.
  • DnaK from Escherichia coli is a model Hsp70.

Purpose of the Study:

  • To investigate the binding orientation of different peptides to the DnaK substrate-binding domain (SBD).
  • To identify the determinants of peptide binding directionality.

Main Methods:

  • Site-directed fluorescence labeling and quenching using bimane and tryptophan.
  • Analysis of fluorescence quenching to determine peptide orientation.
  • Computer modeling of peptide-DnaK(SBD) complexes.

Related Experiment Videos

Main Results:

  • Peptides with N- or C-terminal charged residues (NRLLLTG, NTLLLRG) bound in a forward orientation.
  • Peptides with central proline residues (ELPLVKI, ELPPVKI) showed a strong preference for reverse orientation.
  • Computer modeling suggested steric constraints and hydrogen bonding from central residues dictate orientation.

Conclusions:

  • DnaK can bind substrates in both forward and reverse orientations.
  • Central peptide residues are the primary determinants of binding directionality.
  • This finding advances understanding of chaperone-substrate interactions.