Conversion of B800 Bacteriochlorophyll a to 3-Acetyl Chlorophyll a in the Light-Harvesting Complex 3 by In Situ Oxidation

The spectral features of energy donors and acceptors and the relationship between them in photosynthetic light-harvesting proteins are crucial for photofunctions of these proteins. Engineering energy donors and acceptors in light -harvesting proteins affords the means to increase our under-standing of their photofunctional mechanisms. Herein, we demonstrate the conversion of energy-donating B800 bacterio-chlorophyll (BChl) a to 3-acetyl chlorophyll (AcChl) a in light -harvesting complex 3 (LH3) from Rhodoblastus acidophilus by in situ oxidation with 2,3-dichloro-5,6-dicyano-1,4-benzoquinone. AcChl a in the B800 site exhibited a Qy band that was 111 nm blue-shifted with respect to BChl a in oxidized LH3. The structure of LH3 was barely influenced by the oxidation process, based on circular dichroism spectroscopy and size-exclusion chromatography evidence. In oxidized LH3, AcChl a transferred excitation energy to B820 BChl a, but the rate of excitation energy transfer (EET) was lower than in native LH3. The intracomplex EET in oxidized LH3 was slightly faster than in oxidized light -harvesting complex 2 (LH2). This difference is rationalized by an increase in overlap of the luminescence band of AcChl a with the long tail of the B820 absorption band in oxidized LH3 compared with that of the B850 band in oxidized LH2.