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

Double-mutant cycles: a powerful tool for analyzing protein structure and function

A Horovitz1

  • 1Department of Structural Biology, Weizmann Institute of Science, Rehovot, Israel. csamnon@weizmann.weizmann.ac.il

Folding & Design
|January 1, 1996
PubMed
Summary
This summary is machine-generated.

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

Gynodioecy as a possible populational strategy for increasing reproductive output.

TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik·2013
Same author

How an Ebstein-Barr virus may induce acute fulminant myocarditis in a young immunocompetent adult: a case report.

The West Indian medical journal·2013
Same author

Review: allostery in chaperonins.

Journal of structural biology·2001
Same author

Nested allosteric interactions in the cytoplasmic chaperonin containing TCP-1.

Protein science : a publication of the Protein Society·2001
Same author

In vivo and in vitro function of GroEL mutants with impaired allosteric properties.

The Journal of biological chemistry·2000
Same author

On the relationship between the Hill coefficients for steady-state and transient kinetic data: a criterion for concerted transitions in allosteric proteins.

Bulletin of mathematical biology·2000
Same journal

Fishing for folding nuclei in lattice models and proteins.

Folding & design·1999
Same journal

Folding nucleus: specific or multiple? Insights from lattice models and experiments.

Folding & design·1999
Same journal

Sequential domain refolding of pig muscle 3-phosphoglycerate kinase: kinetic analysis of reactivation.

Folding & design·1999
Same journal

Discrete molecular dynamics studies of the folding of a protein-like model.

Folding & design·1999
Same journal

Knowledge-based structure prediction of MHC class I bound peptides: a study of 23 complexes.

Folding & design·1999
Same journal

How and why phosphotyrosine-containing peptides bind to the SH2 and PTB domains.

Folding & design·1999
See all related articles

Double-mutant cycle analysis reveals coupled residues in proteins by measuring free energy changes. This method quantizes pairwise interactions and characterizes challenging protein structures.

Area of Science:

  • Biochemistry
  • Structural Biology
  • Protein Engineering

Background:

  • Protein function and structure are governed by interactions between residues.
  • Understanding these interactions is crucial for protein engineering and drug design.
  • Traditional methods struggle to characterize certain transient or difficult-to-crystallize protein states.

Purpose of the Study:

  • To introduce and explain the principles of double-mutant cycle analysis.
  • To highlight its utility in quantifying residue coupling and interaction strengths.
  • To demonstrate its application in studying complex or inaccessible protein structures.

Main Methods:

  • Constructing double-mutant cycles involving wild-type, single, and double mutant proteins.
  • Measuring changes in free energy associated with mutations.

Related Experiment Videos

  • Analyzing the additivity or non-additivity of free energy changes to infer residue coupling.

    • Main Results:

    • Identified coupled residues indicate direct or indirect interactions.
    • Quantified the strength of intramolecular and intermolecular pairwise interactions.
    • Enabled characterization of protein structures inaccessible to NMR and X-ray crystallography, including transition states and membrane proteins.

    Conclusions:

    • Double-mutant cycle analysis is a powerful tool for dissecting protein interactions.
    • It provides quantitative insights into residue coupling and cooperativity.
    • The method extends structural and functional characterization to challenging biological systems.