Linear dichroism (LD) spectroscopy is an important technique in the study of the orientation and organization of pigments in the photosynthetic membrane complexes in vivo and in vitro . In this work, the orient...Linear dichroism (LD) spectroscopy is an important technique in the study of the orientation and organization of pigments in the photosynthetic membrane complexes in vivo and in vitro . In this work, the orientation of the pigments in the isolated photosystem Ⅱ (PSⅡ) sub_core reaction center complexes was analyzed and characterized by means of low temperature absorption and LD spectroscopy. The preparations containing different amounts of CP47 isolated from spinach (Spinacia oleracea L.) chloroplast were used in order to investigate the orientation of pigments in the PSⅡ sub_core CP47/D1/D2/Cyt b_559 (CP47/D1/D2) complexes. Chlorophyll a (Chl a) absorbing at 680 nm in CP47/D1/D2/Cyt b_559 complex showed an orientation of the Q y transition parallel to the membrane plane. It is proposed that there are two forms of β_carotene (β_Car) in CP47/D1/D2/Cyt b_559 complex, denoted as β_Car (Ⅰ) and β_Car (Ⅱ), with different orientations, β_Car (Ⅰ) at 470 and 505 nm is roughly parallel to the membrane plane, and β_Car (Ⅱ) at 460 and 490 nm seems to be perpendicular orientation. Upon the photoinhibitory experiment β_Car (Ⅱ) was found to be photosensitive and easily photodamaged. It also showed that the positive LD signal observed at 680 nm was quite complicated. This signal is tentatively attributed to P680 and some Chl a of antenna in CP47 protein based upon our measurements.展开更多
Ultrafast time_resolved fluorescence experiments have been performed with core antennas CP43 and CP47 of PS Ⅱ. Their dynamic fluorescence spectra were obtained with excitation wavelength 514.5 nm. For CP43, the emiss...Ultrafast time_resolved fluorescence experiments have been performed with core antennas CP43 and CP47 of PS Ⅱ. Their dynamic fluorescence spectra were obtained with excitation wavelength 514.5 nm. For CP43, the emission spectrum was found to be from 640 to 780 nm with a peak at ～680 nm and the lifetime of fluorescence was 3.54 ns. For CP47, the emission spectrum was from 630 to 775 nm with a peak at ～691 nm and the fluorescence lifetime was 3.22 ns. The fluorescence emission efficiencies of Chl a in CP43 and CP47 were calculated to be approximately 38.3% and 40.6%, respectively. The energy transfer from β_Car to Chl a in CP43 and CP47 was discussed. The rates of energy transfer from β_Car to Chl a were measured to be about 9.6×10 11 s -1 and 1.3×10 12 s -1 and energy transfer efficiencies 47.5% and 66.5% respectively. The edge_edge distances between β_Car and Chl a in CP43 and CP47 were estimated to be ～0.110 nm and ～0.085 nm respectively.展开更多
The structural and functional alterations within the PSⅡ membrane from phosphatidylcholine reconstitution and Triton X_100 (TX_100) treatment were studied by using Fourier transform_infrared (FT_IR) spectroscopic tec...The structural and functional alterations within the PSⅡ membrane from phosphatidylcholine reconstitution and Triton X_100 (TX_100) treatment were studied by using Fourier transform_infrared (FT_IR) spectroscopic technique and oxygen electrode. Phosphatidylcholine reconstitution showed no significant effect on the protein secondary structures of PSⅡ membrane but an increase of the rate of PSⅡ_mediated oxygen_evolution. The phosphatidylcholine lipids with different length of acyl chains displayed different capabilities to stimulate oxygen_evolution. In contrast, perturbation of the bilayer lipids by TX_100 resulted in obvious changes of the protein secondary structures within the PSⅡ membrane and in the loss of the PSⅡ_mediated oxygen_evolving activity. The results indicate the importance of membrane integrity in maintaining the stability of the photosynthetic membrane proteins.展开更多
基金Commissaire Energie de Atomique de Francethe 9th Five-Year Major Program of the National Natural Science Foundation of China(39890390)+1 种基金the State Key Basic Research Development Plan of China(973)(G1998010100)Innovation of Laboratory of Photosynthests Basic Research,Institute of Botany,The Chinese Acadeny of Sciences
文摘Linear dichroism (LD) spectroscopy is an important technique in the study of the orientation and organization of pigments in the photosynthetic membrane complexes in vivo and in vitro . In this work, the orientation of the pigments in the isolated photosystem Ⅱ (PSⅡ) sub_core reaction center complexes was analyzed and characterized by means of low temperature absorption and LD spectroscopy. The preparations containing different amounts of CP47 isolated from spinach (Spinacia oleracea L.) chloroplast were used in order to investigate the orientation of pigments in the PSⅡ sub_core CP47/D1/D2/Cyt b_559 (CP47/D1/D2) complexes. Chlorophyll a (Chl a) absorbing at 680 nm in CP47/D1/D2/Cyt b_559 complex showed an orientation of the Q y transition parallel to the membrane plane. It is proposed that there are two forms of β_carotene (β_Car) in CP47/D1/D2/Cyt b_559 complex, denoted as β_Car (Ⅰ) and β_Car (Ⅱ), with different orientations, β_Car (Ⅰ) at 470 and 505 nm is roughly parallel to the membrane plane, and β_Car (Ⅱ) at 460 and 490 nm seems to be perpendicular orientation. Upon the photoinhibitory experiment β_Car (Ⅱ) was found to be photosensitive and easily photodamaged. It also showed that the positive LD signal observed at 680 nm was quite complicated. This signal is tentatively attributed to P680 and some Chl a of antenna in CP47 protein based upon our measurements.
文摘Ultrafast time_resolved fluorescence experiments have been performed with core antennas CP43 and CP47 of PS Ⅱ. Their dynamic fluorescence spectra were obtained with excitation wavelength 514.5 nm. For CP43, the emission spectrum was found to be from 640 to 780 nm with a peak at ～680 nm and the lifetime of fluorescence was 3.54 ns. For CP47, the emission spectrum was from 630 to 775 nm with a peak at ～691 nm and the fluorescence lifetime was 3.22 ns. The fluorescence emission efficiencies of Chl a in CP43 and CP47 were calculated to be approximately 38.3% and 40.6%, respectively. The energy transfer from β_Car to Chl a in CP43 and CP47 was discussed. The rates of energy transfer from β_Car to Chl a were measured to be about 9.6×10 11 s -1 and 1.3×10 12 s -1 and energy transfer efficiencies 47.5% and 66.5% respectively. The edge_edge distances between β_Car and Chl a in CP43 and CP47 were estimated to be ～0.110 nm and ～0.085 nm respectively.
文摘The structural and functional alterations within the PSⅡ membrane from phosphatidylcholine reconstitution and Triton X_100 (TX_100) treatment were studied by using Fourier transform_infrared (FT_IR) spectroscopic technique and oxygen electrode. Phosphatidylcholine reconstitution showed no significant effect on the protein secondary structures of PSⅡ membrane but an increase of the rate of PSⅡ_mediated oxygen_evolution. The phosphatidylcholine lipids with different length of acyl chains displayed different capabilities to stimulate oxygen_evolution. In contrast, perturbation of the bilayer lipids by TX_100 resulted in obvious changes of the protein secondary structures within the PSⅡ membrane and in the loss of the PSⅡ_mediated oxygen_evolving activity. The results indicate the importance of membrane integrity in maintaining the stability of the photosynthetic membrane proteins.
