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

In vitro formation of gap junction vesicles.

D A Goodenough

    The Journal of Cell Biology
    |February 1, 1976
    PubMed
    Summary

    Researchers developed a new method to isolate gap junction vesicles using enzyme digestion. This technique preserves the connexon structure and reveals insights into the gap junction protein composition.

    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

    Gap Junction Structures: III. The Effect of Variations in the Isolation Procedure.

    Biophysical journal·2009
    Same author

    Claudin-8 interacts with multi-PDZ domain protein 1 (MUPP1) and reduces paracellular conductance in epithelial cells.

    Cellular and molecular biology (Noisy-le-Grand, France)·2003
    Same author

    Prenatal lens development in connexin43 and connexin50 double knockout mice.

    Investigative ophthalmology & visual science·2001
    Same author

    Molecular cloning, expression analysis, and functional characterization of connexin44.1: a zebrafish lens gap junction protein.

    Developmental dynamics : an official publication of the American Association of Anatomists·2001
    Same author

    Oocyte-granulosa cell heterologous gap junctions are required for the coordination of nuclear and cytoplasmic meiotic competence.

    Developmental biology·2000
    Same author

    Gap junctional communication in the early Xenopus embryo.

    The Journal of cell biology·2000

    Area of Science:

    • Cell biology
    • Biochemistry
    • Structural biology

    Background:

    • Gap junctions are essential for intercellular communication.
    • Understanding their structure and protein components is crucial for cellular function.

    Purpose of the Study:

    • To develop a method for isolating pure gap junction vesicles.
    • To characterize the structural integrity and protein composition of these vesicles.

    Main Methods:

    • Enzyme digestion using trypsin, collagenase, and hyaluronidase.
    • Structural analysis via electron microscopy and buoyant density measurements.
    • Permeability studies using horseradish peroxidase and cytochrome c.

    Main Results:

    • A method yielding purified gap junction vesicles with intact hexagonal connexon lattices was established.
    • Vesicles exhibited smooth, particle-free membrane faces.
    • Approximately 20% of vesicles were impermeable to proteins >12,000 daltons.
    • Evidence suggests a potential nonjunctional contaminating peptide from disulfide reduction of the main protein.

    Conclusions:

    • The isolation method effectively preserves gap junction structure.
    • The primary gap junction protein (connexin) is approximately 18,000 daltons, potentially reducing to a 9,000-dalton peak.
    • Further investigation is needed to determine the peptide composition of the reduced peak.

    Related Experiment Videos