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

Full, reversible copper removal from ascorbate oxidase.

I Savini, L Morpurgo, L Avigliano

    Biochemical and Biophysical Research Communications
    |September 30, 1985
    PubMed
    Summary

    Copper can be effectively rebound to ascorbate oxidase apo-enzyme using a specific Cu(I) complex, especially under anaerobic conditions, restoring enzymatic activity.

    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

    Diet and primary prevention of stroke: Systematic review and dietary recommendations by the ad hoc Working Group of the Italian Society of Human Nutrition.

    Nutrition, metabolism, and cardiovascular diseases : NMCD·2018
    Same author

    Luteolin-7-glucoside inhibits IL-22/STAT3 pathway, reducing proliferation, acanthosis, and inflammation in keratinocytes and in mouse psoriatic model.

    Cell death & disease·2016
    Same author

    Effects of glyphosate on growth rate, metabolic rate and energy reserves of early juvenile crayfish, Cherax quadricarinatus M.

    Bulletin of environmental contamination and toxicology·2014
    Same author

    Skeletal muscle differentiation: role of dehydroepiandrosterone sulfate.

    Hormone and metabolic research = Hormon- und Stoffwechselforschung = Hormones et metabolisme·2011
    Same author

    Human platelets express authentic CB₁ and CB₂ receptors.

    Current neurovascular research·2010
    Same author

    Effects of laser radiation on Rhus vernicifera laccase, Type 2 Cu-depleted laccase, and stellacyanin.

    Journal of inorganic biochemistry·2009

    Area of Science:

    • Biochemistry
    • Enzymology
    • Bioinorganic Chemistry

    Background:

    • Ascorbate oxidase is a copper-containing enzyme crucial for plant defense and cell wall metabolism.
    • Copper ions are essential for the structure and catalytic activity of ascorbate oxidase.
    • Understanding copper reincorporation into apo-ascorbate oxidase is vital for enzyme reconstitution and function studies.

    Purpose of the Study:

    • To investigate the efficient reincorporation of copper into apo-ascorbate oxidase.
    • To compare copper binding and activity recovery under aerobic and anaerobic conditions.
    • To determine the preferred copper binding site in the apo-enzyme.

    Main Methods:

    • Depletion of copper from ascorbate oxidase using anaerobic cyanide treatment.
    • Incubation of apo-ascorbate oxidase with cupric ions and a stable Cu(I) complex ([Cu(I)(thiourea)3]Cl).
    • Quantification of reincorporated copper and measurement of reconstituted enzyme activity.

    Main Results:

    • Complete copper depletion from ascorbate oxidase was achieved with cyanide.
    • Reconstitution with cupric ions showed minimal copper reincorporation.
    • Incubation with a stoichiometric Cu(I) complex resulted in high copper reincorporation (85-90% anaerobically, 70-75% aerobically).
    • Preferential binding of copper to the blue copper site occurred with substoichiometric Cu(I) complex amounts, leading to lower activity recovery than copper reincorporation.

    Conclusions:

    • A stable Cu(I) complex is effective for reconstituting ascorbate oxidase activity.
    • Anaerobic conditions enhance copper reincorporation efficiency.
    • Preferential copper binding at the blue copper site can occur, impacting overall enzymatic activity recovery.

    Related Experiment Videos