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

Exploring local and non-local interactions for protein stability by structural motif engineering.

M Niggemann1, B Steipe

  • 1Genzentrum der Ludwig-Maximilians-Universität, Feodor-Lynen Strasse 25, München, 81377, Germany.

Journal of Molecular Biology
|February 5, 2000
PubMed
Summary

Local and non-local protein interactions contribute equally to stability. Local interactions enhance resistance to denaturation, while non-local interactions boost folding cooperativity, revealing insights into protein engineering.

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

Notch inhibition counteracts Paneth cell death in absence of caspase-8.

Virchows Archiv : an international journal of pathology·2018
Same author

Metal free carboamination of internal alkynes--an easy access to polysubstituted quinolines.

Chemical communications (Cambridge, England)·2016
Same author

Picosecond time-resolved FRET in the fluorescent protein from Discosoma Red (wt-DsRed).

Chemphyschem : a European journal of chemical physics and physical chemistry·2013
Same author

Single-molecule microscopy of the green fluorescent protein using simultaneous two-color excitation.

Chemphyschem : a European journal of chemical physics and physical chemistry·2013
Same author

Nh3D: a reference dataset of non-homologous protein structures.

BMC structural biology·2005
Same author

Kelvin probe force microscopy study on conjugated polymer/fullerene bulk heterojunction organic solar cells.

Nano letters·2005

Area of Science:

  • Protein biochemistry and biophysics
  • Structural biology
  • Computational biology

Background:

  • Understanding protein thermodynamic stability is crucial for protein engineering.
  • The relative contributions of local and non-local interactions to protein stability remain incompletely understood.
  • Motif engineering and sequence shuffling offer potential avenues to dissect these contributions.

Purpose of the Study:

  • To experimentally determine the relative contributions of local and non-local interactions to protein thermodynamic stability.
  • To assess the impact of replacing non-local interaction-dominated segments with locally interacting motifs.
  • To quantify the role of local interactions in protein folding cooperativity and resistance to denaturation.

Main Methods:

Related Experiment Videos

  • Devised an experimental approach combining motif engineering and sequence shuffling.
  • Identified and replaced candidate chain segments in an immunoglobulin V(L) domain with locally interacting motifs using consensus sequences.
  • Analyzed folding transitions and cooperativity of engineered proteins and shuffled sequences.

    • Main Results:

    • All nine engineered replacements systematically reduced folding cooperativity.
    • Sequence shuffling in host-guest experiments indicated the presence of designed interactions in the native state.
    • Local and non-local interactions were found to contribute approximately equally to protein stability.

    Conclusions:

    • Local interactions primarily increase resistance to denaturation, while non-local interactions enhance folding cooperativity.
    • Consensus sequence engineering is a valuable tool for protein structure design, but global stability requires consideration of non-local interactions.
    • Analysis solely based on local sequence propensities is insufficient for accurately predicting protein structure and stability.