采用聚丙烯酰胺凝胶电泳(PAGE)和蔗糖密度梯度超速离心方法分离了假根羽藻(Bryopsis corticulans)的色素-蛋白复合物,并对其特性进行分析。结果表明:采用PAGE分离得到7条色素-蛋白复合物带,分别是CP I a1、CP I a2、CP I、LHCP1、LHCP2...采用聚丙烯酰胺凝胶电泳(PAGE)和蔗糖密度梯度超速离心方法分离了假根羽藻(Bryopsis corticulans)的色素-蛋白复合物,并对其特性进行分析。结果表明:采用PAGE分离得到7条色素-蛋白复合物带,分别是CP I a1、CP I a2、CP I、LHCP1、LHCP2、CPa、LHCP3+3,和2条游离色素(free pigment,FP)FCa、FC。用改进的不连续蔗糖密度梯度离心法分离到五条带。区带Ⅰ是FP;区带Ⅱ主要是小分子量的PSⅡ捕光复合物LHCP3+3;区带Ⅲ以PSⅡ捕光复合物的聚集体LHCP1为主,区带Ⅱ和Ⅲ的吸收光谱中除了Chla外,还含有大量的Chlb和管藻黄素,是管藻黄素-Chla/b-蛋白质复合物;区带Ⅳ在PAGE中只显示一条带,光谱中有Chlb吸收肩峰,含有66和56kDa两种多肽,是较小的PSⅠ复合物CPⅠa。展开更多
A light-harvesting chlorophyll a/b-protein complex (LHCP) was isolated directly from thylakoid mem-branes of marine green alga, Bryopsis corticulans, by two consecutive runs of liquid chromatography. The trimeric form...A light-harvesting chlorophyll a/b-protein complex (LHCP) was isolated directly from thylakoid mem-branes of marine green alga, Bryopsis corticulans, by two consecutive runs of liquid chromatography. The trimeric form of the light-harvesting complex has been obtained by sucrose gradient ultracentrifugation. The result of SDS- PAGE shows that the light-harvesting complex is composed of at least five apoproteins in which a protein with apparent molecular weight of about 31 kD was never found in the ma-jor light-harvesting complex (LHC Ⅱ) from higher plants. The isolated Bryopsis corticulans light-harvesting complex contains a specific carotenoid, siphonaxanthin, as well as chlorophyll (Chl) a, Chl b, neoxanthin and violaxanthin. Si-phonaxanthin which is present in the light-harvesting sipho-naxanthin-chlorophyll a/b-protein complex of Bryopsis corticulans is responsible for enhanced absorption in the blue-green region (530 nm). Efficient energy transfer from both siphonaxanthin and Chl b to Chl a in Bryopsis corticu-lans LHCP, which has similar absorption and fluorescence emission spectra to those of the lutein-chlorophyll a/b-protein of higher plants, proved that molecular arrangement of the light-harvesting pigments was highly ordered in the Bryopsis corticulans LHCP. The siphonaxanthin-chlorophyll a/b-pro- teins allow enhanced absorption of blue-green light, the pre-dominant light available in deep ocean waters or shaded subtidal marine habitats.展开更多
Steady-state and time-resolved fluo- rescence spectroscopies have been used to study the excited state properties of Chl a in different ag- gregation forms of light-harvesting complex II (LHC II) from an intertidal gr...Steady-state and time-resolved fluo- rescence spectroscopies have been used to study the excited state properties of Chl a in different ag- gregation forms of light-harvesting complex II (LHC II) from an intertidal green alga, Bryopsis corticulans, i.e. LHC II monomer, trimer and oligomer. When either Chl a or Chl b was selectively excited, the observed decrease in Chl a fluorescence in the oligomer is proved to be caused mainly by the fast fluorescence quenching among Chl a molecules, rather than by the decrease in Chl b-to-Chl a singlet excitation transfer efficiency. Analyses of the picosecond time-resolved fluorescence kinetics identified two exponential de- cay components in all of the three forms of LHC II: a longer-lived component (4.1―4.7 ns) originating from fluorescence emission of Chl a, and a shorter-lived one (135―540 ps) from the rapid equilibration of singlet excitation among Chl a molecules. The time constant of excitation equilibration is 135 ps in oli- gomer, 520 ps in trimer and 540 ps in monomer. These results imply that LHC II in oligomer form is inherently able to quench Chl a excitation, a mecha- nism which may be related to the photoprotection of PS II via changing the degree of LHC II aggregation in Bryopsis corticulans.展开更多
文摘采用聚丙烯酰胺凝胶电泳(PAGE)和蔗糖密度梯度超速离心方法分离了假根羽藻(Bryopsis corticulans)的色素-蛋白复合物,并对其特性进行分析。结果表明:采用PAGE分离得到7条色素-蛋白复合物带,分别是CP I a1、CP I a2、CP I、LHCP1、LHCP2、CPa、LHCP3+3,和2条游离色素(free pigment,FP)FCa、FC。用改进的不连续蔗糖密度梯度离心法分离到五条带。区带Ⅰ是FP;区带Ⅱ主要是小分子量的PSⅡ捕光复合物LHCP3+3;区带Ⅲ以PSⅡ捕光复合物的聚集体LHCP1为主,区带Ⅱ和Ⅲ的吸收光谱中除了Chla外,还含有大量的Chlb和管藻黄素,是管藻黄素-Chla/b-蛋白质复合物;区带Ⅳ在PAGE中只显示一条带,光谱中有Chlb吸收肩峰,含有66和56kDa两种多肽,是较小的PSⅠ复合物CPⅠa。
文摘A light-harvesting chlorophyll a/b-protein complex (LHCP) was isolated directly from thylakoid mem-branes of marine green alga, Bryopsis corticulans, by two consecutive runs of liquid chromatography. The trimeric form of the light-harvesting complex has been obtained by sucrose gradient ultracentrifugation. The result of SDS- PAGE shows that the light-harvesting complex is composed of at least five apoproteins in which a protein with apparent molecular weight of about 31 kD was never found in the ma-jor light-harvesting complex (LHC Ⅱ) from higher plants. The isolated Bryopsis corticulans light-harvesting complex contains a specific carotenoid, siphonaxanthin, as well as chlorophyll (Chl) a, Chl b, neoxanthin and violaxanthin. Si-phonaxanthin which is present in the light-harvesting sipho-naxanthin-chlorophyll a/b-protein complex of Bryopsis corticulans is responsible for enhanced absorption in the blue-green region (530 nm). Efficient energy transfer from both siphonaxanthin and Chl b to Chl a in Bryopsis corticu-lans LHCP, which has similar absorption and fluorescence emission spectra to those of the lutein-chlorophyll a/b-protein of higher plants, proved that molecular arrangement of the light-harvesting pigments was highly ordered in the Bryopsis corticulans LHCP. The siphonaxanthin-chlorophyll a/b-pro- teins allow enhanced absorption of blue-green light, the pre-dominant light available in deep ocean waters or shaded subtidal marine habitats.
基金This work was supported by the National Natural Science Foundation of China(Grant No.20273077)the Knowledge Innovation Program of Chinese Academy of Sciences(Grant No.KSCX2-SW-130).
文摘Steady-state and time-resolved fluo- rescence spectroscopies have been used to study the excited state properties of Chl a in different ag- gregation forms of light-harvesting complex II (LHC II) from an intertidal green alga, Bryopsis corticulans, i.e. LHC II monomer, trimer and oligomer. When either Chl a or Chl b was selectively excited, the observed decrease in Chl a fluorescence in the oligomer is proved to be caused mainly by the fast fluorescence quenching among Chl a molecules, rather than by the decrease in Chl b-to-Chl a singlet excitation transfer efficiency. Analyses of the picosecond time-resolved fluorescence kinetics identified two exponential de- cay components in all of the three forms of LHC II: a longer-lived component (4.1―4.7 ns) originating from fluorescence emission of Chl a, and a shorter-lived one (135―540 ps) from the rapid equilibration of singlet excitation among Chl a molecules. The time constant of excitation equilibration is 135 ps in oli- gomer, 520 ps in trimer and 540 ps in monomer. These results imply that LHC II in oligomer form is inherently able to quench Chl a excitation, a mecha- nism which may be related to the photoprotection of PS II via changing the degree of LHC II aggregation in Bryopsis corticulans.