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Updated: Feb 7, 2026

Quantitative FRET Förster Resonance Energy Transfer Analysis for SENP1 Protease Kinetics Determination
Published on: February 21, 2013
Excitation energy transfer kinetics and efficiency in phototrophic green sulfur bacteria
Nikki Cecil M Magdaong1, Dariusz M Niedzwiedzki2, Rafael G Saer3
1Department of Biology, Washington University in St. Louis, St. Louis, MO 63130, USA; Department of Chemistry, Washington University in St. Louis, St. Louis, MO 63130, USA; Photosynthetic Antenna Research Center, Washington University in St. Louis, St. Louis, MO 63130, USA.
Abstract:
A series of spectroscopic measurements were performed on membrane fractions and detergent-solubilized complexes from the green sulfur bacterium (GSB) Chlorobaculum (Cba.) tepidum. The excitation migration through the entire GSB photosynthetic apparatus cannot be observed upon excitation of membranes in the chlorosome region at 77 K. In order to observe energy transfer from the Fenna-Matthews-Olson (FMO) protein to the reaction center (RC), FMO was directly excited at ~800 nm in transient absorption experiments. However, interpretation of the results is complicated by the spectral overlap between FMO and the RC. The availability of the Y16F FMO mutant, whose absorption spectrum is drastically different from that of the WT, has enabled the selection of spectral regions where either only FMO or the RC contributes. The application of a directed kinetic modeling approach, or target analysis, revealed the various decay and energy transfer pathways within the pigment-protein complexes. The calculated FMO-to-RC excitation energy transfer efficiencies are approximately 25% and 48% for the Y16F and WT samples, respectively.
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