English (United Kingdom)

https://curated-unify.zendy.io/wp-json/zendy-region/v1/featured_content/oa?rat=en

https://curated-unify.zendy.io/wp-json/zendy-region/v1/highlighted_journal/

Zendy Open

Presents the pdf analysis as premium feature

PDF Analysis

Insights

Presents the summarisation as premium feature

Summarisation

Presents the keyphrase highlighting as premium feature

Keyphrase Highlighting

Presents the zaia usage as premium feature

ZAIA

Zendy Tools

Zendy Plus

Presents the access of premium content as premium feature

Premium Content

Sedimentation velocity analytical ultracentrifugation with fluorescencedetection has emerged as a powerful method for the study of interacting systemsof macromolecules. It combines picomolar sensitivity with high hydrodynamicresolution, and can be carried out with photoswitchable fluorophores formulti-component discrimination, to determine the stoichiometry, affinity, andshape of macromolecular complexes with dissociation equilibrium constants frompicomolar to micromolar. A popular approach for data interpretation is thedetermination of the binding affinity by isotherms of weight-averagesedimentation coefficients, sw. A prevailing dogma in sedimentation analysis isthat the weight-average sedimentation coefficient from the transport methodcorresponds to the signal- and population-weighted average of all species. Weshow that this does not always hold true for systems that exhibit significantsignal changes with complex formation - properties that may be readilyencountered in practice, e.g., from a change in fluorescence quantum yield.Coupled transport in the reaction boundary of rapidly reversible systems canmake significant contributions to the observed migration in a way that cannotbe accounted for in the standard population-based average. Effective particletheory provides a simple physical picture for the reaction-coupled migrationprocess. On this basis we develop a more general binding model that convergesto the well-known form of sw with constant signals, but can accountsimultaneously for hydrodynamic co-transport in the presence of changes influorescence quantum yield. We believe this will be useful when studyinginteracting systems exhibiting fluorescence quenching, enhancement or Forsterresonance energy transfer with transport methods.

Sedimentation of Reversibly Interacting Macromolecules with Changes in Fluorescence Quantum Yield