FRET-based Stoichiometry Measurements of Protein Complexes

TY - JOUR

T1 - FRET-based Stoichiometry Measurements of Protein Complexes

AU - Mattiroli, Francesca

AU - Gu, Yajie

AU - Luger, Karolin

PY - 2018/2/5

Y1 - 2018/2/5

N2 - 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.

AB - 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.

U2 - 10.21769/bioprotoc.2713

DO - 10.21769/bioprotoc.2713

M3 - Article

C2 - 29644254

SN - 2331-8325

VL - 7

JO - Bio-Protocol

JF - Bio-Protocol

IS - 3

ER -