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

Mapping protein-protein interactions by affinity-directed mass spectrometry

Y Zhao1, T W Muir, S B Kent

  • 1Rockefeller University, New York, NY 10021, USA.

Proceedings of the National Academy of Sciences of the United States of America
|April 30, 1996
PubMed
Summary

Researchers developed affinity-directed mass spectrometry for precise protein interaction site identification. This method rapidly maps binding epitopes on growth factors and can analyze other molecular interactions.

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

Access to site-specific Fc-cRGD peptide conjugates through streamlined expressed protein ligation.

Organic & biomolecular chemistry·2016
Same author

Origin and removal of adducts (molecular mass = 98 u) attached to peptide and protein ions in electrospray ionization mass spectra.

Journal of the American Society for Mass Spectrometry·2013
Same author

Matrix-assisted laser desorption mass spectrometry of biotransformation products of dynorphin a in vitro.

Journal of the American Society for Mass Spectrometry·2013
Same author

Collision induced decomposition of peptides. Choice of collision parameters.

Journal of the American Society for Mass Spectrometry·2013
Same author

Detection and correction of interference in SRM analysis.

Methods (San Diego, Calif.)·2013
Same author

Native chemical ligation of polypeptides.

Current protocols in protein science·2008

Area of Science:

  • Biochemistry
  • Analytical Chemistry
  • Molecular Biology

Background:

  • Identifying protein interaction sites is crucial for understanding biological processes.
  • Existing methods can be time-consuming or lack precision.

Purpose of the Study:

  • To demonstrate a precise and rapid method for identifying protein interaction sites.
  • To map the binding epitope of a growth factor with a monoclonal antibody.

Main Methods:

  • Developed and applied affinity-directed mass spectrometry.
  • Utilized proteolytic digestion and analysis of synthetic peptide ladders.
  • Direct mass spectrometric readout from protein complexes.

Main Results:

  • Successfully mapped the approximate location of a continuous binding epitope on a growth factor.

Related Experiment Videos

  • Precisely determined the boundaries of the binding epitope.
  • Demonstrated the method's applicability to ligand-receptor and protein-oligonucleotide interactions.

    • Conclusions:

    • Affinity-directed mass spectrometry offers a precise and rapid approach for mapping molecular interactions.
    • The method is versatile and applicable to various molecule-molecule contacts.
    • Enables efficient characterization of binding epitopes and interaction sites.