In vivo analysis of the 2-Cys peroxiredoxin oligomeric state by two-step FRET

摘要: Fluorescence resonance energy transfer (FRET) analysis in biological systems has reached broad application along with the fast improvement of fluorescent proteins. This work shows advancement commonly used single-step FRET between two fluorophores to a two-step three vivo. In addition CFP and YFP DsRed derivative mCherry was genetically fused frame coding region plastidic 2-Cys peroxiredoxin co-expressed plant cells resulting detectable radiationless from via mCherry. The use control constructs such as fluorophore pairs CFP, mCherry, but also YFP:mCherry:CFP REACh:mCherry:CFP allowed for generation reference matrix calculations. occurrence proves that obligate dimers assemble higher mass oligomers presumably decamers finding together previous reports on structural dynamics functional switching might indicate conformation linked redox-signalling function Prx. Although different fusion proteins had be imported by chloroplast significant within complex. proof oligomerisation vivo, results demonstrate large potential method investigating tripartite protein interactions subcellular compartments general cell biology.