Electrochromic responses of carotenoid absorbance bands in purified light-harvesting complexes from Rhodobacter capsulatus reconstituted into liposomes

Abstract

Light-Harvesting Complexes I and II (LHI and LHII) were extracted from chromatophores of Rhodobacter capsulatus, purified in Triton X-100 and reconstituted into phospholipid vesicles. Application of membrane potentials (K + diffusion potentials) to LHII proteoliposomes led to absorbance changes in the carotenoid bands which were spectrally similar to those in chromatophores. These (electrochromic) absorbance changes were linear with the applied membrane potential between −107 mV and +105 mV. The data were consistent with the existence of two forms of carotenoid in LHII. One form, comprising 2 3 of the total and with a long wavelength absorbance maximum at 510 nm, was not significantly affected by membrane potential. The other component, comprising 1 3 of the total and with a long wavelength absorbance maximum at 516.5 nm, was shifted by approx. 1.6 nm to the red by a membrane potential of 105 mV. Reduction of the B800 bacteriochlorophyll in LHII with NaBH 4 before reconstitution did not affect the absorbance spectrum of the carotenoids and it did not affect their response to applied membrane potentials in proteoliposomes. Although the electrochromically-sensitive carotenoids might be associated with B800, interactions with the bacteriochlophyll are perhaps not the cause of the polarisation of the carotenoid that is responsible for the linearity of the response. The carotenoids in reconstituted LHI complexes were not detectably electrochromic. The electrochromic absorbance changes of carotenoids in LHII could be useful for membrane potential measurement in liposomes containing ion-translocating proteins.