Recent advances in the proteomic field have allowed high-throughput experiments to be conducted on chloroplast samples. Many proteomic investigations have focused on either whole chloroplast or sub-plastidial fraction...Recent advances in the proteomic field have allowed high-throughput experiments to be conducted on chloroplast samples. Many proteomic investigations have focused on either whole chloroplast or sub-plastidial fractions. To date, the Plant Protein Database （PPDB, Sun et al., 2009） presents the most exhaustive chloroplast proteome available online. However, the accurate localization of many proteins that were identified in different sub-plastidial compartments remains hypothetical. Ferro et al. （2009） went a step further into the knowledge of Arabidopsis thaliana chloroplast proteins with regards to their accurate localization within the chloroplast by using a semi-quantitative proteomic approach known as spectral counting. Their proteomic strategy was based on the accurate mass and time tags （AMT） database approach and they built up AT_CHLORO, a comprehensive chloroplast proteome database with sub-plastidial localization and curated information on envelope proteins. Comparing these two extensive databases, we focus here on about 100 enzymes involved in the synthesis of chloroplast-specific isoprenoids. Well known pathways （i.e. compartmentation of the methyl erythritol phosphate biosynthetic pathway, of tetrapyrroles and chlorophyll biosynthesis and breakdown within chloroplasts） validate the spectral counting-based strategy. The same strategy was then used to identify the precise localization of the biosynthesis of carotenoids and prenylquinones within chloroplasts （i.e. in envelope membranes, stroma, and/or thylakoids） that remains unclear until now.展开更多
This paper aimed to evaluate the effects of different concentrations of cadmium on growth rates, photosynthetic pigments, photosynthetic performance, biochemical parameters and structure of chloroplasts in G. domingen...This paper aimed to evaluate the effects of different concentrations of cadmium on growth rates, photosynthetic pigments, photosynthetic performance, biochemical parameters and structure of chloroplasts in G. domingensis. To accomplish this, apical segments of G. domingensis were cultivated with different concentrations of cadmium, ranging from 100 to 300 μM, over a period of 16 days, and were processed for transmission electron microscopy analysis. The plants exposed to cadmium showed chloroplast alteration, especially degeneration of thylakoids and a decrease in the concentration of photosynthetic pigments, such as chlorophyll a and phycobiliproteins. However, the volume of plastoglobuli increased. As a defense mechanism, the plants treated with cadmium showed an increase in glutathione reductase activity. These results agree with the decreased photosynthetic performance and relative electron transport rate observed after exposure of algae to cadmium. Taken together, these findings strongly indicate that cadmium negatively affects the ultrastructure and metabolism of the agarophyte G. domingensis, thus posing a threat to the economic vitality of this red macroalga.展开更多
The bottom-up construction of self-powered artificial cells is significant to understand the energy supply and metabolism of nature cells.Here,we demonstrate an efficient manner to build thylakoid-containing artificia...The bottom-up construction of self-powered artificial cells is significant to understand the energy supply and metabolism of nature cells.Here,we demonstrate an efficient manner to build thylakoid-containing artificial cells,which continuously convert light energy into chemical energy to supply adenosine 5'-triphosphate(ATP)under light illumination.The production of ATP supplies energy to promote the biological enzyme cascade reactions,where glucose is transformed into glucose 6-phosphate(G6P)under the catalysis of hexokinase(HK).G6P was further converted to gluconolactone 6-phosphate(PG)in the presence of 6-phosphate dehydrogenase(G6PDH),meanwhile NADP^(+) was converted to nicotinamide adenine dinucleotide phosphate(NADPH).The self-powered artificial cells were demonstrated to generate ATP and NADPH successively,which provided a way for building more complicated artificial cells.展开更多
A phosphetase that hydrolyses phosphate monoesters has been Isolated from wheat thylakold membranes. Biochemical properties and inhibition kinetics of the phosphatase were Investigated using several Ions, organlc solv...A phosphetase that hydrolyses phosphate monoesters has been Isolated from wheat thylakold membranes. Biochemical properties and inhibition kinetics of the phosphatase were Investigated using several Ions, organlc solvents, and Inhlbltors. Wheat （Trltlcum aestivum L. cv. PH82-2-2） thylakold membrane phosphatase activity was activated by Mg^2＋, Ca^2＋, and Fe^2＋ and was inhibited by Mn^2＋ and Cu^2＋. For example, enzyme activity was acUvated 34.81% by 2 mmol/l. Mg^2＋, but was Inhibited 22.3% and 8.5% by 2 and 1 mmol/L Cu^2＋, respectively. Methanol, ethanol and glycol were all able to activate enzyme activity. Enzyme activity was activated 58.5%, 48.2%, and 8.7% by 40% ethanol, methanol and glycol, respectively. From these results, It can be seen that the degree of actlvetlon of the phosphetase was greatest for ethanol and the type of acUvatlon was uncompetltlve. Moreover, the activity of the thylakold membrane phosphetase was Inhibited by molybdate, vanadete, phosphate, and fluoride and the type of Inhibition produced by these elements was uncompetltlve, non-competitive, competltlve and mixed, respectively.展开更多
Unicellular cyanobacteria Synechocystis 6803 were fixed using high-pressure freezing （HPF） and freeze substitution without any chemical cross-linkers. Immunoelectron microscopy of these cells showed that five sequen...Unicellular cyanobacteria Synechocystis 6803 were fixed using high-pressure freezing （HPF） and freeze substitution without any chemical cross-linkers. Immunoelectron microscopy of these cells showed that five sequential enzymes of the Calvin cycle （phosphoriboisomerase, phosphoribulokinase, ribulose-1,5-bisphosphate carboxylase/oxygenase （RuBisCO）, 3-phosphoglyceratekinase and glyceraldehyde-3-phosphate dehydrogenase） and the catalytic portion of the chloroplast H^＋-ATP synthase （CF1） are located adjacent to the thylakoid membranes. Cell-free extracts of Synechocystis were processed by ultracentrifugation to isolate thylakoid fractions sedimenting at 40 000, 90 000, and 150 000 g. Among these, the 150 000-g fraction showed the highest linked activity of the above five sequential Calvin cycle enzymes and also the highest coordinated activity of light and dark reactions as assessed by ribose-5-phosphate （R-5-P） ＋ADP dependent CO2 fixation. Immunogold labeling of this membrane fraction confirmed the presence of the above five enzymes as well as the catalytic portion of the CF1 ATP synthase. Notably, the protein A-gold labeling of the thylakoids was observed without use of chemical cross-linkers and in spite of the normal washing steps used during standard immunolabeling. The results showed that soluble Calvin cycle enzymes might be organized along the thylakoid membranes.展开更多
Proanthocyanidins are formed in the chlorophyllous organs of Tracheophyta from a redifferentiation of chloroplasts involving the thylakoidal membrane and lumen. With the purpose to help researchers of concerned discip...Proanthocyanidins are formed in the chlorophyllous organs of Tracheophyta from a redifferentiation of chloroplasts involving the thylakoidal membrane and lumen. With the purpose to help researchers of concerned disciplines to identify such chloroplasts, we described herein the morphologies of functional and redifferentiating chloroplasts in various members of Tracheophyta. The most obvious sign of redifferentiation is a tremendous swelling of the chloroplast which turns obese. De novo genesis of osmiophilic materials is also characteristic, either as single dots attached to the inner face of the swollen thylakoidal membrane which will yield the tannosomes, or as pearl necklace-shaped structures protruding into the lumen;this last formation can be viewed as a giant tannosome forming finally stromal chlorotannic accretions. Whatever their mode of formation is, tannosomes are expulsed from the chloroplast as shuttles.展开更多
文摘Recent advances in the proteomic field have allowed high-throughput experiments to be conducted on chloroplast samples. Many proteomic investigations have focused on either whole chloroplast or sub-plastidial fractions. To date, the Plant Protein Database （PPDB, Sun et al., 2009） presents the most exhaustive chloroplast proteome available online. However, the accurate localization of many proteins that were identified in different sub-plastidial compartments remains hypothetical. Ferro et al. （2009） went a step further into the knowledge of Arabidopsis thaliana chloroplast proteins with regards to their accurate localization within the chloroplast by using a semi-quantitative proteomic approach known as spectral counting. Their proteomic strategy was based on the accurate mass and time tags （AMT） database approach and they built up AT_CHLORO, a comprehensive chloroplast proteome database with sub-plastidial localization and curated information on envelope proteins. Comparing these two extensive databases, we focus here on about 100 enzymes involved in the synthesis of chloroplast-specific isoprenoids. Well known pathways （i.e. compartmentation of the methyl erythritol phosphate biosynthetic pathway, of tetrapyrroles and chlorophyll biosynthesis and breakdown within chloroplasts） validate the spectral counting-based strategy. The same strategy was then used to identify the precise localization of the biosynthesis of carotenoids and prenylquinones within chloroplasts （i.e. in envelope membranes, stroma, and/or thylakoids） that remains unclear until now.
文摘This paper aimed to evaluate the effects of different concentrations of cadmium on growth rates, photosynthetic pigments, photosynthetic performance, biochemical parameters and structure of chloroplasts in G. domingensis. To accomplish this, apical segments of G. domingensis were cultivated with different concentrations of cadmium, ranging from 100 to 300 μM, over a period of 16 days, and were processed for transmission electron microscopy analysis. The plants exposed to cadmium showed chloroplast alteration, especially degeneration of thylakoids and a decrease in the concentration of photosynthetic pigments, such as chlorophyll a and phycobiliproteins. However, the volume of plastoglobuli increased. As a defense mechanism, the plants treated with cadmium showed an increase in glutathione reductase activity. These results agree with the decreased photosynthetic performance and relative electron transport rate observed after exposure of algae to cadmium. Taken together, these findings strongly indicate that cadmium negatively affects the ultrastructure and metabolism of the agarophyte G. domingensis, thus posing a threat to the economic vitality of this red macroalga.
基金supported by the National Natural Science Foundation of China(Grant Nos.21929401,,22174031,22111540252,21773050)the Fundamental Research Funds forthe Central Universities(HIT.OCEF.2021026)+1 种基金the Heilongjiang Touyan Team(HITTY-20190034)the Natural Science Foundation of Heilongjiang Province(ZD2022B001).
文摘The bottom-up construction of self-powered artificial cells is significant to understand the energy supply and metabolism of nature cells.Here,we demonstrate an efficient manner to build thylakoid-containing artificial cells,which continuously convert light energy into chemical energy to supply adenosine 5'-triphosphate(ATP)under light illumination.The production of ATP supplies energy to promote the biological enzyme cascade reactions,where glucose is transformed into glucose 6-phosphate(G6P)under the catalysis of hexokinase(HK).G6P was further converted to gluconolactone 6-phosphate(PG)in the presence of 6-phosphate dehydrogenase(G6PDH),meanwhile NADP^(+) was converted to nicotinamide adenine dinucleotide phosphate(NADPH).The self-powered artificial cells were demonstrated to generate ATP and NADPH successively,which provided a way for building more complicated artificial cells.
文摘A phosphetase that hydrolyses phosphate monoesters has been Isolated from wheat thylakold membranes. Biochemical properties and inhibition kinetics of the phosphatase were Investigated using several Ions, organlc solvents, and Inhlbltors. Wheat （Trltlcum aestivum L. cv. PH82-2-2） thylakold membrane phosphatase activity was activated by Mg^2＋, Ca^2＋, and Fe^2＋ and was inhibited by Mn^2＋ and Cu^2＋. For example, enzyme activity was acUvated 34.81% by 2 mmol/l. Mg^2＋, but was Inhibited 22.3% and 8.5% by 2 and 1 mmol/L Cu^2＋, respectively. Methanol, ethanol and glycol were all able to activate enzyme activity. Enzyme activity was activated 58.5%, 48.2%, and 8.7% by 40% ethanol, methanol and glycol, respectively. From these results, It can be seen that the degree of actlvetlon of the phosphetase was greatest for ethanol and the type of acUvatlon was uncompetltlve. Moreover, the activity of the thylakold membrane phosphetase was Inhibited by molybdate, vanadete, phosphate, and fluoride and the type of Inhibition produced by these elements was uncompetltlve, non-competitive, competltlve and mixed, respectively.
文摘Unicellular cyanobacteria Synechocystis 6803 were fixed using high-pressure freezing （HPF） and freeze substitution without any chemical cross-linkers. Immunoelectron microscopy of these cells showed that five sequential enzymes of the Calvin cycle （phosphoriboisomerase, phosphoribulokinase, ribulose-1,5-bisphosphate carboxylase/oxygenase （RuBisCO）, 3-phosphoglyceratekinase and glyceraldehyde-3-phosphate dehydrogenase） and the catalytic portion of the chloroplast H^＋-ATP synthase （CF1） are located adjacent to the thylakoid membranes. Cell-free extracts of Synechocystis were processed by ultracentrifugation to isolate thylakoid fractions sedimenting at 40 000, 90 000, and 150 000 g. Among these, the 150 000-g fraction showed the highest linked activity of the above five sequential Calvin cycle enzymes and also the highest coordinated activity of light and dark reactions as assessed by ribose-5-phosphate （R-5-P） ＋ADP dependent CO2 fixation. Immunogold labeling of this membrane fraction confirmed the presence of the above five enzymes as well as the catalytic portion of the CF1 ATP synthase. Notably, the protein A-gold labeling of the thylakoids was observed without use of chemical cross-linkers and in spite of the normal washing steps used during standard immunolabeling. The results showed that soluble Calvin cycle enzymes might be organized along the thylakoid membranes.
文摘Proanthocyanidins are formed in the chlorophyllous organs of Tracheophyta from a redifferentiation of chloroplasts involving the thylakoidal membrane and lumen. With the purpose to help researchers of concerned disciplines to identify such chloroplasts, we described herein the morphologies of functional and redifferentiating chloroplasts in various members of Tracheophyta. The most obvious sign of redifferentiation is a tremendous swelling of the chloroplast which turns obese. De novo genesis of osmiophilic materials is also characteristic, either as single dots attached to the inner face of the swollen thylakoidal membrane which will yield the tannosomes, or as pearl necklace-shaped structures protruding into the lumen;this last formation can be viewed as a giant tannosome forming finally stromal chlorotannic accretions. Whatever their mode of formation is, tannosomes are expulsed from the chloroplast as shuttles.
