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

Detergent removal during membrane reconstitution

T M Allen, A Y Romans, H Kercret

    Biochimica Et Biophysica Acta
    |September 18, 1980
    PubMed
    Summary
    This summary is machine-generated.

    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

    Aetiology of Dental Caries.

    The American journal of dental science·2019
    Same author

    Lipid nanoparticle delivery systems for siRNA-based therapeutics.

    Drug delivery and translational research·2015
    Same author

    Within-session responding as a function of post-session feedings.

    Behavioural processes·2014
    Same author

    Association of the family environment with behavioural and cognitive outcomes in children with chromosome 22q11.2 deletion syndrome.

    Journal of intellectual disability research : JIDR·2013
    Same author

    Randomized multicenter clinical trial of myofascial physical therapy in women with interstitial cystitis/painful bladder syndrome and pelvic floor tenderness.

    The Journal of urology·2012
    Same author

    Low-power resonant laser ablation of copper.

    Applied optics·2010
    Same journal

    Cumulative Contents.

    Biochimica et biophysica acta·2020
    Same journal

    Molecular Basis of Disease Cumulative Contents.

    Biochimica et biophysica acta·2020
    Same journal

    General Subjects Cumulative Contents.

    Biochimica et biophysica acta·2020
    Same journal

    Erratum to 'on the role of exchangeable hydrogen bonds for the kinetics of P680<sup>+·</sup> Q<sub>A</sub> <sup>-·</sup> formation and P680<sup>+·</sup> Pheo<sup>-·</sup> recombination in photosystem II' [Biochim. Biophys. Acta 1276 (1996) 35-44].

    Biochimica et biophysica acta·2019
    Same journal

    Oligomeric state of the light-harvesting complexes B800-850 and B875 from purple bacterium Rubrivivax gelatinosus in detergent solution.

    Biochimica et biophysica acta·2019
    Same journal

    Regulation of pigment content and enzyme activity in the cyanobacterium Nostoc sp. Mac grown in continuous light, a light-dark photoperiod, or darkness.

    Biochimica et biophysica acta·2019
    See all related articles

    Detergent removal for membrane protein reconstitution is most efficient using gel filtration, significantly outperforming dialysis methods for various detergents and ethanol. This method also aids in quantifying protein-lipid interactions.

    Area of Science:

    • Biochemistry
    • Membrane Protein Research
    • Lipid Bilayer Studies

    Background:

    • Membrane protein reconstitution is crucial for studying protein function.
    • Efficient removal of detergents and ethanol used in reconstitution is essential for accurate results.
    • Existing methods like dialysis and gel filtration vary in their effectiveness.

    Purpose of the Study:

    • To compare the efficiency of detergent and ethanol removal using different reconstitution methods.
    • To evaluate the effectiveness of gel filtration versus dialysis for removing specific detergents (sodium deoxycholate, cholic acid, Triton X-100) and ethanol.
    • To explore the use of gel filtration for quantifying protein-lipid affinities.

    Main Methods:

    • Ethanol injection-dialysis

    Related Experiment Videos

  • Detergent dialysis
  • Detergent-gel filtration using Sephadex G-200/G-50 and Biobeads SM-2
  • Competition assays using glycophorin T(is) peptide, sodium deoxycholate, and phospholipids

    • Main Results:

    • Ethanol injection-dialysis retained significant ethanol (approx. 70/1000 phospholipid molecules).
    • Detergent removal by dialysis was detergent-dependent, with residual sodium deoxycholate (approx. 7/1000 phospholipid molecules) even after extensive dialysis.
    • Gel filtration demonstrated superior detergent removal efficiency compared to dialysis, retaining fewer detergent molecules (e.g., 10 deoxycholate/100 phospholipid molecules after one passage).
    • Ethanol removal was less efficient with gel filtration than extensive dialysis.
    • Biobeads SM-2 and Sephadex gel filtration showed comparable Triton X-100 removal.
    • Preliminary studies indicated a lipid affinity order for glycophorin T(is) peptide: phosphatidylinositol > phosphatidylcholine > phosphatidylserine.

    Conclusions:

    • Gel filtration is a more efficient method for removing detergents like sodium deoxycholate, cholic acid, and Triton X-100 during membrane protein reconstitution compared to dialysis.
    • While effective for detergents, gel filtration is less efficient for ethanol removal than prolonged dialysis.
    • Gel filtration techniques show promise for quantitative assessment of protein-lipid interactions.