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

Design of protein function leaps by directed domain interface evolution.

Jin Huang1, Akiko Koide, Koki Makabe

  • 1Department of Biochemistry and Molecular Biology, University of Chicago, 929 East 57th Street, Chicago, IL 60637, USA.

Proceedings of the National Academy of Sciences of the United States of America
|May 1, 2008
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

Promiscuous RNA binding by WDR5 remodels the KMT2A (MLL1) histone methyltransferase complex to an inactive state.

bioRxiv : the preprint server for biology·2026
Same author

Protein Engineering-Enabled Cryo-EM Investigation of Small GTPases.

Journal of molecular biology·2026
Same author

The integrated stress response promotes immune evasion through lipocalin 2.

Nature·2026
Same author

Engineered antibodies that stabilize drug-modified KRAS<sup>G12C</sup> neoantigens enable selective and potent cross-HLA immunotherapy.

Nature communications·2025
Same author

Engineering antibody-drug conjugates targeting an adhesion GPCR, CD97.

Proceedings of the National Academy of Sciences of the United States of America·2025
Same author

Generation of actionable, cancer-specific neoantigens from KRAS(G12C) with adagrasib.

Proceedings of the National Academy of Sciences of the United States of America·2025
Same journal

The TaMYB55-TaSnRK1α1-TabZIP9 module confers heat stress tolerance in wheat.

Proceedings of the National Academy of Sciences of the United States of America·2026
Same journal

Superstatistics approach to turbulent circulation fluctuations.

Proceedings of the National Academy of Sciences of the United States of America·2026
Same journal

A molecular timescale for evolution of cobamide biosynthesis.

Proceedings of the National Academy of Sciences of the United States of America·2026
Same journal

Pierre Chambon, a pioneer of molecular biology and gene regulation in eukaryotes.

Proceedings of the National Academy of Sciences of the United States of America·2026
Same journal

Granulosa cell glycogen fuels the avascular corpus luteum.

Proceedings of the National Academy of Sciences of the United States of America·2026
Same journal

Synthetic essentiality of TRAIL/TNFSF10 in VHL-deficient renal cell carcinoma.

Proceedings of the National Academy of Sciences of the United States of America·2026
See all related articles

Researchers created novel protein functions by combining protein domains, significantly boosting binding affinity and specificity. This protein engineering approach yields highly effective affinity reagents, outperforming antibodies in some applications.

Area of Science:

  • Protein engineering
  • Structural biology
  • Molecular evolution

Background:

  • Many sophisticated proteins utilize multiple domains with active sites at domain interfaces.
  • Gene recombination creating new domain interfaces is a proposed mechanism for major functional leaps in proteins.

Purpose of the Study:

  • To demonstrate that complex protein functions can be generated by combining unrelated domains.
  • To develop a new protein engineering strategy for creating high-affinity reagents.

Main Methods:

  • Joining a low-affinity peptide-binding domain with an inert domain.
  • Employing directed evolution to optimize the domain interface.
  • Characterizing the resulting protein structure and function using X-ray crystallography.

Related Experiment Videos

Main Results:

  • Achieved >500-fold increase in affinity and >2,000-fold increase in specificity compared to single domains.
  • Developed "affinity clamps" with clamshell architecture and enhanced binding surfaces.
  • Demonstrated single-nanomolar dissociation constants, outperforming monoclonal antibodies in immunochemical assays.

Conclusions:

  • Established a method for evolving sophisticated protein functions from simpler domains via interface engineering.
  • Introduced a novel protein engineering concept for designing targeted, high-affinity reagents.
  • Highlighted the potential for discovering numerous new protein functions through directed domain interface evolution.