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FRET-based Stoichiometry Measurements of Protein Complexes in vitro.

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 | 2018 Feb

REPOSITORIES: biostudies-literature

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FRET-based Stoichiometry Measurements of Protein Complexes <i>in vitro</i>.

Mattiroli Francesca F Gu Yajie Y Luger Karolin K

Bio-protocol 20180201 3

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 <i>in vitro</i>. 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 ...[more]

PMID: 29644254

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