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

Phosphorylase kinase conformers. Detection by proteases.

M R Trempe, G M Carlson

    The Journal of Biological Chemistry
    |March 25, 1987
    PubMed
    Summary

    Protease studies reveal that phosphorylase kinase activation, through phosphorylation or cation binding, alters the beta subunit

    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

    The regulatory alpha subunit of phosphorylase kinase may directly participate in the binding of glycogen phosphorylase.

    Biochemistry. Biokhimiia·2002
    Same author

    Direct visualization of the calmodulin subunit of phosphorylase kinase via electron microscopy following subunit exchange.

    Journal of structural biology·2001
    Same author

    Self-association of the alpha subunit of phosphorylase kinase as determined by two-hybrid screening.

    The Journal of biological chemistry·1999
    Same author

    Structural features contributing to complex formation between glycogen phosphorylase and phosphorylase kinase.

    Biochemistry·1999
    Same author

    Mg2+ induces conformational changes in the catalytic subunit of phosphorylase kinase, whether by itself or as part of the holoenzyme complex.

    Journal of protein chemistry·1999
    Same author

    Activators of phosphorylase kinase alter the cross-linking of its catalytic subunit to the C-terminal one-sixth of its regulatory alpha subunit.

    Biochemistry·1999

    Area of Science:

    • Biochemistry
    • Enzymology
    • Protein Structure and Function

    Background:

    • Phosphorylase kinase is a key enzyme in glycogen metabolism.
    • Understanding its conformational changes is crucial for elucidating its regulation.

    Purpose of the Study:

    • To investigate structural and conformational changes in phosphorylase kinase using protease digestion.
    • To differentiate between activated and nonactivated enzyme conformers.

    Main Methods:

    • Protease digestion (trypsin, chymotrypsin, endoproteinase Arg C) of nonphosphorylated and phosphorylated phosphorylase kinase.
    • Analysis of subunit degradation rates.
    • Effect of activating/inhibitory cations (Mg2+, Ca2+, Mn2+) and allosteric activator (ADP) on proteolysis.

    Main Results:

    • The alpha subunit is rapidly degraded by most proteases.
    • The beta subunit's susceptibility to trypsin differs significantly between nonactivated and activated phosphorylase kinase.
    • Phosphorylation, Mg2+, Ca2+, and ADP protect the beta subunit from tryptic hydrolysis, indicating a conformational change.

    Conclusions:

    • Activation of phosphorylase kinase by various mechanisms induces a conformational change in the beta subunit.
    • Trypsin serves as a reliable probe for detecting these effector-induced conformational changes.
    • These findings align with previous studies using chemical cross-linkers.

    Related Experiment Videos