A lipid_depleted cytochrome b 6f (Cyt b 6f) preparation was obtained from spinach (Spinacia oleracea L.) chloroplasts. Upon reconstitution of this preparation with the membrane lipids purified from spinach thylakoid...A lipid_depleted cytochrome b 6f (Cyt b 6f) preparation was obtained from spinach (Spinacia oleracea L.) chloroplasts. Upon reconstitution of this preparation with the membrane lipids purified from spinach thylakoid, the effects of different membrane lipids on the electron transfer activity were studied. The results show that the electron transfer activity of Cyt b 6f is obviously stimulated to different extents, respectively, by monogalactosyldiacylglycerol (MGDG), digalactosyldiacylglycerol (DGDG), phosphatidylcholine (PC), phosphatidylglycerol (PG) and sulfoquinovosyldiacylglycerol (SQDG), and that the extents of stimulation may be closely related to the charge of the membrane lipids. The stimulation of non_charged lipids (MGDG, DGDG) and neutrally_charged lipid (PC) was high with a maximum enhancement of 89%, 75% and 77%, respectively; but the stimulation of two kinds of negatively_charged lipid (PG and SQDG) was relatively low with a maximum enhancement of 43% and 26%, respectively.展开更多
The singlet excited state lifetime of the chlorophyll a (Chl a) in cytochrome b6f (Cyt b6f) complex was reported to be shorter than that of free Chl a in methanol, but the value was different for Cyt b6f com-plexes fr...The singlet excited state lifetime of the chlorophyll a (Chl a) in cytochrome b6f (Cyt b6f) complex was reported to be shorter than that of free Chl a in methanol, but the value was different for Cyt b6f com-plexes from different sources (~200 and ~600 ps are the two measured results). The present study demonstrated that the singlet excited state lifetime is associated with the detergents n-dodecyl-β-D- maltoside (DDM) and n-octyl-β-D-glucopyranoside (β-OG), but has nothing to do with the different sources of Cyt b6f complexes. Compared with the Cyt b6f dissolved in β-OG, the Cyt b6f in DDM had a lower fluorescence yield, a lower photodegradation rate of Chl a, and a shorter lifetime of Chl a excited state. In short, the singlet excited state lifetime, ~200 ps, of the Chl a in Cyt b6f complex in DDM is closer to the true in vivo.展开更多
The cytochrome b_(6f)(Cyt b_(6f))complex is a multisubunit protein complex in chloroplast thylakoid membranes required for photosynthetic electron transport.Here we report the isolation and characterization of the new...The cytochrome b_(6f)(Cyt b_(6f))complex is a multisubunit protein complex in chloroplast thylakoid membranes required for photosynthetic electron transport.Here we report the isolation and characterization of the new tiny albino 1(nta1)mutant in Arabidopsis,which has severe defects in Cyt b_(6f) accumulation and chloroplast development.Gene cloning revealed that the nta1 phenotype was caused by disruption of a single nuclear gene,NTA1,which encodes an integral thylakoid membrane protein conserved across green algae and plants.Overexpression of NTA1 completely rescued the nta1 phenotype,and knockout of NTA1 in wild-type plants recapitulated the mutant phenotype.Loss of NTA1 function severely impaired the accumulation of multiprotein complexes related to photosynthesis in thylakoid membranes,particularly the components of Cyt b_(6f).NTA1 was shown to directly interact with four subunits(Cyt b6/PetB,PetD,PetG,and PetN)of Cyt b_(6f) through the DUF1279 domain and C-terminal sequence to mediate their assembly.Taken together,our results identify NTA1 as a new and key regulator of chloroplast development that plays essential roles in assembly of the Cyt b_(6f) complex by interacting with multiple Cyt b_(6f) subunits.展开更多
An apoptotic cell-free system containing cytosol and nuclei from normally cultured tobacco sus-pension cells was used to show that a spinach chloroplast preparation can induce apoptosis in nuclei, evi-denced by DNA el...An apoptotic cell-free system containing cytosol and nuclei from normally cultured tobacco sus-pension cells was used to show that a spinach chloroplast preparation can induce apoptosis in nuclei, evi-denced by DNA electrophoresis and fluorescence microscopy observations. Further study showed that the chloroplast preparation or its pellet (thylakoid membrane) after hypoosmotic or supersonic treatment still ex-hibited the apoptosis-inducing activity, but the supernatant had no effect, which indicates that the apoptosis-inducing effector in the chloroplast preparation is water-insoluble. The induction of apoptosis by chloroplast preparation could be attenuated by Ac-DEVD-CHO, the specific inhibitor of Caspase-3, implying involve-ment of a Caspase-3-like protease during the process. Furthermore, extensive apoptosis in nuclei was in-duced by cytochrome b6/f on the thylakoid membrane, indicating that this important cytochrome complex may have an important role in the chloroplast-related apoptotic pathway.展开更多
基金The State Key Basic Research and Development Plan(G1998010100)Innovative Foundation of Laboratory of Photosynthesis Basic Research Insitute of Botany,The Chinese Academy of Sciences
文摘A lipid_depleted cytochrome b 6f (Cyt b 6f) preparation was obtained from spinach (Spinacia oleracea L.) chloroplasts. Upon reconstitution of this preparation with the membrane lipids purified from spinach thylakoid, the effects of different membrane lipids on the electron transfer activity were studied. The results show that the electron transfer activity of Cyt b 6f is obviously stimulated to different extents, respectively, by monogalactosyldiacylglycerol (MGDG), digalactosyldiacylglycerol (DGDG), phosphatidylcholine (PC), phosphatidylglycerol (PG) and sulfoquinovosyldiacylglycerol (SQDG), and that the extents of stimulation may be closely related to the charge of the membrane lipids. The stimulation of non_charged lipids (MGDG, DGDG) and neutrally_charged lipid (PC) was high with a maximum enhancement of 89%, 75% and 77%, respectively; but the stimulation of two kinds of negatively_charged lipid (PG and SQDG) was relatively low with a maximum enhancement of 43% and 26%, respectively.
基金Supported by the National Natural Science Foundation of China (Grant Nos.39890390 and 20673144)
文摘The singlet excited state lifetime of the chlorophyll a (Chl a) in cytochrome b6f (Cyt b6f) complex was reported to be shorter than that of free Chl a in methanol, but the value was different for Cyt b6f com-plexes from different sources (~200 and ~600 ps are the two measured results). The present study demonstrated that the singlet excited state lifetime is associated with the detergents n-dodecyl-β-D- maltoside (DDM) and n-octyl-β-D-glucopyranoside (β-OG), but has nothing to do with the different sources of Cyt b6f complexes. Compared with the Cyt b6f dissolved in β-OG, the Cyt b6f in DDM had a lower fluorescence yield, a lower photodegradation rate of Chl a, and a shorter lifetime of Chl a excited state. In short, the singlet excited state lifetime, ~200 ps, of the Chl a in Cyt b6f complex in DDM is closer to the true in vivo.
基金supported by the General Research Fund(CUHK codes 14121915,14148916,and 14104521)the Area of Excellence Scheme(AoE/M-403/16 and AoE/M-05/12)of the Research Grants Council(RGC)of Hong Kong+1 种基金the National Natural Science Foundation of China(NSFC)-RGC Joint Scheme(N_CUHK452/17)direct grants from the Chinese University of Hong Kong(CUHK).
文摘The cytochrome b_(6f)(Cyt b_(6f))complex is a multisubunit protein complex in chloroplast thylakoid membranes required for photosynthetic electron transport.Here we report the isolation and characterization of the new tiny albino 1(nta1)mutant in Arabidopsis,which has severe defects in Cyt b_(6f) accumulation and chloroplast development.Gene cloning revealed that the nta1 phenotype was caused by disruption of a single nuclear gene,NTA1,which encodes an integral thylakoid membrane protein conserved across green algae and plants.Overexpression of NTA1 completely rescued the nta1 phenotype,and knockout of NTA1 in wild-type plants recapitulated the mutant phenotype.Loss of NTA1 function severely impaired the accumulation of multiprotein complexes related to photosynthesis in thylakoid membranes,particularly the components of Cyt b_(6f).NTA1 was shown to directly interact with four subunits(Cyt b6/PetB,PetD,PetG,and PetN)of Cyt b_(6f) through the DUF1279 domain and C-terminal sequence to mediate their assembly.Taken together,our results identify NTA1 as a new and key regulator of chloroplast development that plays essential roles in assembly of the Cyt b_(6f) complex by interacting with multiple Cyt b_(6f) subunits.
基金the National Natural Science Foundation of China (No. 39970072)
文摘An apoptotic cell-free system containing cytosol and nuclei from normally cultured tobacco sus-pension cells was used to show that a spinach chloroplast preparation can induce apoptosis in nuclei, evi-denced by DNA electrophoresis and fluorescence microscopy observations. Further study showed that the chloroplast preparation or its pellet (thylakoid membrane) after hypoosmotic or supersonic treatment still ex-hibited the apoptosis-inducing activity, but the supernatant had no effect, which indicates that the apoptosis-inducing effector in the chloroplast preparation is water-insoluble. The induction of apoptosis by chloroplast preparation could be attenuated by Ac-DEVD-CHO, the specific inhibitor of Caspase-3, implying involve-ment of a Caspase-3-like protease during the process. Furthermore, extensive apoptosis in nuclei was in-duced by cytochrome b6/f on the thylakoid membrane, indicating that this important cytochrome complex may have an important role in the chloroplast-related apoptotic pathway.
