FRET-based Stoichiometry Measurements of Protein Complexes in vitro.
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ABSTRACT: For a complete understanding of biochemical reactions, information on complex stoichiometry is essential. However, measuring stoichiometry is experimentally challenging. Our lab has developed a FRET-based assay to study protein complex stoichiometry in vitro. This assay, also known as Job plot, is set up as a continuous variation of the molar ratio between the two species, kept at constant total concentration. The FRET (Fluorescence Resonance Energy Transfer) between the two fluorescently-labeled proteins is measured and the stoichiometry is inferred from the sample with highest FRET signal. This approach allows us to assess complex stoichiometry in solution.
SUBMITTER: Mattiroli F
PROVIDER: S-EPMC5891122 | biostudies-literature |
REPOSITORIES: biostudies-literature
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