Cytochrome b_559 in photosystem Ⅱ reaction center was purified from spinach ( Spinacia oleracea L.) and rice ( Oryza sativa L.) by a rapid and simple procedure. Their low temperature fluorescence emission and e...Cytochrome b_559 in photosystem Ⅱ reaction center was purified from spinach ( Spinacia oleracea L.) and rice ( Oryza sativa L.) by a rapid and simple procedure. Their low temperature fluorescence emission and excitation spectra, ultraviolet fluorescence spectra and absolute absorption spectra were presented. The author's purification methods, which enhanced the yield of pure protein and shorted the time for isolation, have several advantages: 1. use of oxygen_evolving PSⅡ core complexes as the starting material in order to avoid disturbing from other cytochromes; 2. isocratic elution of cytochrome b_559 from a DEAE_Sephacel column for eliminating the impurity and yielding the protein in pure state; 3. a simple column procedure for removal of excess Triton X_100. Purified cytochromes b_559 from these species have similar optical spectra and mobility during gel electrophoresis under native conditions. From the results of novel electrophoresis (Tricine_SDS_PAGE), cytochrome b_559 from both spinach and rice reveal two polypeptide bands (apparent molecular weight 9 kD and 4 kD, respectively). By measuring of 77 K fluorescence spectra, it was shown that for the purified cytochrome b_559 there were two excitation peaks at 439 nm and 413 nm, and two emission peaks at 563 nm and 668 nm. This is the first indication that Cyt b_559 is able to emit fluorescence and also transfer excited electrons to chlorophyll. By the use of ultraviolet fluorescence spectra, it was demonstrated for the first time that the location of Trp residue could be in the hydrophobic transmembrane region of cytochrome b_559.展开更多
The influence of digalactosyldiacylglycerol (DGDG), one of the photosynthetic membrane lipids, on heat inactivation of the process of oxygen evolution has been studied in vitro in photosystem Ⅱ(PS Ⅱ) core complex. I...The influence of digalactosyldiacylglycerol (DGDG), one of the photosynthetic membrane lipids, on heat inactivation of the process of oxygen evolution has been studied in vitro in photosystem Ⅱ(PS Ⅱ) core complex. It was found that the temperature of semi-inactivation of oxygen evolution in the complex increased from 40.0 to about 43.0℃ in the presence of DGDG with 5-min heat treatment in the dark. Furthermore, when PS Ⅱ core complex was incubated for 5 min at 45.0℃, the oxygen evolution in the complex was completely lost, whilst the DGDG-complexed PS Ⅱ core complex still retained a 16% of activity (100% for 25.0℃). In addition, a 1-h incubation at 38.0℃ inactivated absolutely the oxygen evolution for the PS Ⅱ core complex. By contrast, there remained about 20% of activity (zero time for 100%) for the complex in the presence of DGDG under the same condition. These results indicate a new role of DGDG in the protection of PS Ⅱ core complex against the deleterious effects of temperature. It was展开更多
PSⅡ core antenna complexes, CP43 and CP47, were purified from spinach (Spinacia oleracea L.) by DEAE Fractogel TSK 650S anion exchange chromatography. Their normal temperature (298 K) resonance Raman spectra were...PSⅡ core antenna complexes, CP43 and CP47, were purified from spinach (Spinacia oleracea L.) by DEAE Fractogel TSK 650S anion exchange chromatography. Their normal temperature (298 K) resonance Raman spectra were measured. The results suggest that all β carotenoids bound to CP43 and CP47 are in all trans configuration and likely in twisted conformations.展开更多
Photodamage of some pigments in the isolated photosystem Ⅱ (PS Ⅱ) reaction center D1/D2/Cyt b559 complex from spinach has been investigated by means of high performance liquid chromatography. The light induced damag...Photodamage of some pigments in the isolated photosystem Ⅱ (PS Ⅱ) reaction center D1/D2/Cyt b559 complex from spinach has been investigated by means of high performance liquid chromatography. The light induced damage of pheophytin a (pheo a) in the complex was observed for the first time. The content of pheo a decreased about 47% by illumination, suggesting only one of the two pheo a molecules in the PSⅡ reaction center complex was damaged. No damage of β carotene was found.展开更多
Photodamage of pheophytin a (pheo a) in the isolated photosystem Ⅱ (PSⅡ) reaction center D1/D2/Cyt b559 complex from spinach has been investigated by high performance liquid chromatographic method in detail. The res...Photodamage of pheophytin a (pheo a) in the isolated photosystem Ⅱ (PSⅡ) reaction center D1/D2/Cyt b559 complex from spinach has been investigated by high performance liquid chromatographic method in detail. The results showed that: (1) There is one pheo a molecule which is not associated with the primary photochemistry in the PSⅡ reaction center complex. It may be considered that there are two different electron transfer branches in the PSⅡ reaction center just as in the purple bacterium photosynthetic reaction center. (2) The damaged pheo a may be attributed to the one bonding to the D2 protein comparing the D2 subunit in the PSⅡ reaction center with M subunit in the purple bacterium photosynthetic reaction center. (3) A possible arrangement model of redox cofactors in the PSⅡ reaction center was proposed based on our experiment.展开更多
光系统Ⅱ(photosystemⅡ,PSⅡ)是位于植物、藻类和蓝细菌等放氧光合生物类囊体膜上的重要超分子复合物,它可通过捕获光能用于激发反应中心的电荷分离并驱动电子传递过程,在常温常压下可将水分子裂解产生氧气和质子.植物光系统Ⅱ的外周...光系统Ⅱ(photosystemⅡ,PSⅡ)是位于植物、藻类和蓝细菌等放氧光合生物类囊体膜上的重要超分子复合物,它可通过捕获光能用于激发反应中心的电荷分离并驱动电子传递过程,在常温常压下可将水分子裂解产生氧气和质子.植物光系统Ⅱ的外周存在主要和次要捕光复合物Ⅱ(major and minor light-harvesting complexⅡ,LHCⅡ),它们负责吸收光能并向光系统Ⅱ传递激发能,并且还参与非光化学淬灭和状态转换相关的捕光调节过程.近年来,围绕光系统Ⅱ和LHCⅡ的结构生物学研究取得了一系列重要进展,本文总结了PSⅡ、LHCⅡ和二者共同组成的PSII-LHCII超级复合物的结构生物学研究历程以及最新进展,并对该领域的未来研究方向做出展望.展开更多
Light and heat induced denaturation of CP47, the core antenna complex of photosystem Ⅱ purified from spinach, were investigated using absorption and circular dichroism spectra. Light caused the destruction of chlorop...Light and heat induced denaturation of CP47, the core antenna complex of photosystem Ⅱ purified from spinach, were investigated using absorption and circular dichroism spectra. Light caused the destruction of chlorophyll a and excitonic interaction of chlorophyll a in CP47, while the protein secondary structure was not apparently changed. Heat induced the destruction of protein secondary structure and excitonic interaction of chlorophyll a, but the chlorophyll a molecule was not damaged. The results suggest that both the chlorophyll a molecular structure and the protein native conformation are necessary for excitonic interaction of chlorophyll a and the energy transfer function of the chlorophyll a binding protein.展开更多
文摘Cytochrome b_559 in photosystem Ⅱ reaction center was purified from spinach ( Spinacia oleracea L.) and rice ( Oryza sativa L.) by a rapid and simple procedure. Their low temperature fluorescence emission and excitation spectra, ultraviolet fluorescence spectra and absolute absorption spectra were presented. The author's purification methods, which enhanced the yield of pure protein and shorted the time for isolation, have several advantages: 1. use of oxygen_evolving PSⅡ core complexes as the starting material in order to avoid disturbing from other cytochromes; 2. isocratic elution of cytochrome b_559 from a DEAE_Sephacel column for eliminating the impurity and yielding the protein in pure state; 3. a simple column procedure for removal of excess Triton X_100. Purified cytochromes b_559 from these species have similar optical spectra and mobility during gel electrophoresis under native conditions. From the results of novel electrophoresis (Tricine_SDS_PAGE), cytochrome b_559 from both spinach and rice reveal two polypeptide bands (apparent molecular weight 9 kD and 4 kD, respectively). By measuring of 77 K fluorescence spectra, it was shown that for the purified cytochrome b_559 there were two excitation peaks at 439 nm and 413 nm, and two emission peaks at 563 nm and 668 nm. This is the first indication that Cyt b_559 is able to emit fluorescence and also transfer excited electrons to chlorophyll. By the use of ultraviolet fluorescence spectra, it was demonstrated for the first time that the location of Trp residue could be in the hydrophobic transmembrane region of cytochrome b_559.
基金This work was supported by the National Natural Science Foundation of China (Grant No. 39890390)the State Basic Research Development Plan of China (Grant No. G1998010100)the Innovative Foundation of Laboratory of Photosynthesis Basic Research, In
文摘The influence of digalactosyldiacylglycerol (DGDG), one of the photosynthetic membrane lipids, on heat inactivation of the process of oxygen evolution has been studied in vitro in photosystem Ⅱ(PS Ⅱ) core complex. It was found that the temperature of semi-inactivation of oxygen evolution in the complex increased from 40.0 to about 43.0℃ in the presence of DGDG with 5-min heat treatment in the dark. Furthermore, when PS Ⅱ core complex was incubated for 5 min at 45.0℃, the oxygen evolution in the complex was completely lost, whilst the DGDG-complexed PS Ⅱ core complex still retained a 16% of activity (100% for 25.0℃). In addition, a 1-h incubation at 38.0℃ inactivated absolutely the oxygen evolution for the PS Ⅱ core complex. By contrast, there remained about 20% of activity (zero time for 100%) for the complex in the presence of DGDG under the same condition. These results indicate a new role of DGDG in the protection of PS Ⅱ core complex against the deleterious effects of temperature. It was
文摘PSⅡ core antenna complexes, CP43 and CP47, were purified from spinach (Spinacia oleracea L.) by DEAE Fractogel TSK 650S anion exchange chromatography. Their normal temperature (298 K) resonance Raman spectra were measured. The results suggest that all β carotenoids bound to CP43 and CP47 are in all trans configuration and likely in twisted conformations.
文摘Photodamage of some pigments in the isolated photosystem Ⅱ (PS Ⅱ) reaction center D1/D2/Cyt b559 complex from spinach has been investigated by means of high performance liquid chromatography. The light induced damage of pheophytin a (pheo a) in the complex was observed for the first time. The content of pheo a decreased about 47% by illumination, suggesting only one of the two pheo a molecules in the PSⅡ reaction center complex was damaged. No damage of β carotene was found.
文摘Photodamage of pheophytin a (pheo a) in the isolated photosystem Ⅱ (PSⅡ) reaction center D1/D2/Cyt b559 complex from spinach has been investigated by high performance liquid chromatographic method in detail. The results showed that: (1) There is one pheo a molecule which is not associated with the primary photochemistry in the PSⅡ reaction center complex. It may be considered that there are two different electron transfer branches in the PSⅡ reaction center just as in the purple bacterium photosynthetic reaction center. (2) The damaged pheo a may be attributed to the one bonding to the D2 protein comparing the D2 subunit in the PSⅡ reaction center with M subunit in the purple bacterium photosynthetic reaction center. (3) A possible arrangement model of redox cofactors in the PSⅡ reaction center was proposed based on our experiment.
文摘光系统Ⅱ(photosystemⅡ,PSⅡ)是位于植物、藻类和蓝细菌等放氧光合生物类囊体膜上的重要超分子复合物,它可通过捕获光能用于激发反应中心的电荷分离并驱动电子传递过程,在常温常压下可将水分子裂解产生氧气和质子.植物光系统Ⅱ的外周存在主要和次要捕光复合物Ⅱ(major and minor light-harvesting complexⅡ,LHCⅡ),它们负责吸收光能并向光系统Ⅱ传递激发能,并且还参与非光化学淬灭和状态转换相关的捕光调节过程.近年来,围绕光系统Ⅱ和LHCⅡ的结构生物学研究取得了一系列重要进展,本文总结了PSⅡ、LHCⅡ和二者共同组成的PSII-LHCII超级复合物的结构生物学研究历程以及最新进展,并对该领域的未来研究方向做出展望.
基金the National Natural Science Foundation of China !(No .39890 390 )
文摘Light and heat induced denaturation of CP47, the core antenna complex of photosystem Ⅱ purified from spinach, were investigated using absorption and circular dichroism spectra. Light caused the destruction of chlorophyll a and excitonic interaction of chlorophyll a in CP47, while the protein secondary structure was not apparently changed. Heat induced the destruction of protein secondary structure and excitonic interaction of chlorophyll a, but the chlorophyll a molecule was not damaged. The results suggest that both the chlorophyll a molecular structure and the protein native conformation are necessary for excitonic interaction of chlorophyll a and the energy transfer function of the chlorophyll a binding protein.
