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

New fluorogenic, photoactivable, heterobifunctional crosslinking thiol reagents.

T Ueno, S Hikita, D Muno

    Analytical Biochemistry
    |July 1, 1984
    PubMed
    Summary

    Newly developed photoactivable crosslinking reagents (ACM) enable sensitive detection of protein crosslinking. These reagents specifically bind to sulfhydryl groups and become fluorescent upon photoactivation, facilitating analysis of protein 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

    Corneal advanced glycation end products increase in patients with proliferative diabetic retinopathy.

    Diabetes care·2001
    Same author

    Somatic mutation of mitochondrial DNA in cancerous and noncancerous liver tissue in individuals with hepatocellular carcinoma.

    Cancer research·2001
    Same author

    [Evans syndrome].

    Ryoikibetsu shokogun shirizu·2001
    Same author

    Inflammatory cytokines modulate eotaxin release by human lung fibroblast cell line.

    Experimental lung research·2001
    Same author

    Immunoliposome-mediated gene transfer into cultured myotubes.

    The Tohoku journal of experimental medicine·2001
    Same author

    Facilitatory and neurotoxic effects of intravitreal ornithine on the electroretinographic responses of albino rats.

    Ophthalmic research·2001

    Area of Science:

    • Biochemistry
    • Molecular Biology
    • Protein Chemistry

    Background:

    • Protein crosslinking is crucial for studying protein structure and interactions.
    • Existing crosslinking methods may lack specificity or sensitive detection capabilities.
    • Photoactivable reagents offer advantages in controlled crosslinking and visualization.

    Purpose of the Study:

    • To synthesize and characterize novel photoactivable, heterobifunctional crosslinking reagents (ACM).
    • To investigate the application of ACM reagents for specific protein modification and crosslinking.
    • To evaluate the utility of ACM reagents for sensitive detection of crosslinked protein products.

    Main Methods:

    • Synthesis of photoactivable heterobifunctional crosslinking reagents (ACM) with maleimide and azide groups.

    Related Experiment Videos

  • Specific binding of ACM reagents to protein sulfhydryl residues.
  • Photoactivation of the azide group for intra- or intermolecular crosslinking.
  • Detection of crosslinked products using fluorometry and SDS-PAGE.

    • Main Results:

    • ACM reagents demonstrated specific binding to protein sulfhydryl groups.
    • Photoactivation of ACM led to efficient intra- and intermolecular protein crosslinking.
    • The coumarin moiety of ACM became highly fluorescent post-photolysis, enabling sensitive detection.
    • Application to rabbit muscle aldolase showed extensive subunit crosslinking, with up to 25% crosslinked to dimers.

    Conclusions:

    • ACM reagents are effective tools for specific protein crosslinking and sensitive detection.
    • The photoactivable and fluorescent properties of ACM enhance the study of protein interactions.
    • ACM reagents provide a valuable method for analyzing protein subunit associations and structural dynamics.