• Title, Summary, Keyword: Thylakoid membrane

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The Effects of Surfactants on the Biosynthesis of Galactolipid and the Composition of Fatty Acids in Chloroplast Envelope rind Thylakoid Membrane of Chlorella ellipsoidea

  • Choe, Eun-A;Cheong, Gyeong-Suk;Lee, Cheong-Sam
    • Animal cells and systems
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    • v.2 no.3
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    • pp.341-349
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    • 1998
  • To analyze the effects of surfactants on the biosynthesis of galactolipid and the composition of fatty acids, the chloroplast envelope and thylakoid membrane were cultivated in medium treated with anionic surfactants, such as linear alkylbenzene sulfonate (0.002%, LAS), a-olefin sulfonate (O.01%, AOS), and sodium lauryl ether sulfate (0.08%, SLES), respectively. During the cultivation, the chloroplast envelope and thylakoid membrane were isolated from the cells collected at the early and middle phase of the culture and the contents of their fatty acid composition were compared with the control. When treated with surfactants, the contents of total lipid MDGD methylesters, and DGDG methylesters decreased significantly when compared with the control. It was also confirmed that more unsaturated fatty acids were involved in the biosynthesis of galactolipid. The fatty acids utilized in the biosynthesis of MGDG were in the chloroplast envelope and in the control, and linoleic acid in LAS, linolenic acid and oleic acid in AOS, and linolenic acid and oleic acid in SLES. The fatty acids in the biosynthesis of DGDG were linolenic acid and oleic acid in the control linolenic acid and stearic acid in LAS, oleic acid and linolenic acid in AOS, oleic acid and linolenic acid in SLES. In the thylakoid membrane, the major fatty acids in the biosynthesis of MGDG were linolenic acid and oleic acid in the control, oleic acid and linolenic acid in LAS, linolenic acid and linoleic acid in AOS, linolenic acid and palmitoleic acid in SLES. The fatty acids in the biosynthesis of DGDG were linolenic acid and oleic acid in the control, oleic acid and linolenic acid in LAS, linolenic acid and linoleic acid in AOS, palmitoleic acid and oleic acid in SLES.

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Changes in Protein Synthesis Induced by Chilling in Tomato Chloroplasts

  • Kim, Won-Il;Jung, Goo-Bok;Kim, Min-Kyeong;Park, Kwang-Lai;Yun, Sun-Gang
    • Korean Journal of Environmental Agriculture
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    • v.20 no.5
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    • pp.310-316
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    • 2001
  • To find out the effect of low temperature on the regulation of tomato chloroplast genes, the optimization of the system in chloroplast protein synthesis and the identification of the changes in chloroplast protein synthesis induced by chilling were studied. Incorporation reaction occurred rapidly at the first 30 minutes and was constantly maintained after 60 minutes. A broad optimal temperature on protein synthesis was found around 20 to $30^{\circ}C$. No difference was shown in the chloroplast protein synthesis under high light intensity (1600 ${\mu}E/m^2/s$) as well as under low light intensity (400 ${\mu}E/m^2/s$) even darkness. $K^+$, $Mg^{++}$ and ATP at an optimal concentration act as an activator, while DTT, chloramphenicol, cycloheximide, $Ca^{++}$ and inorganic phosphate act as an inhibitor in the chloroplast protein synthesis. Synthesis of 15, 55 and 60 kd chloroplast encoded stromal proteins and 18, 24, 33 and 55 kd chloroplast encoded thylakoid membrane proteins were reduced by chilling, while 17 kd chloroplast encoded stromal protein and 16 kd chloroplast encoded thylakoid membrane protein was induced by chilling. It was expected that the 55 kd stromal protein would be the large subunit of rubisco and the 33 kd thylakoid membrane protein would be the D1 protein which was drastically reduced by chilling.

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Characteristics of Photosynthetic Electron Transport Activity in Isolated Chloroplast of Korean Ginseng and Radish (인삼과 무 엽록체의 광합성 전자전달 활성)

  • 김갑식
    • Journal of Plant Biology
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    • v.33 no.2
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    • pp.111-118
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    • 1990
  • In order to characterize the chloroplasts of Korean ginseng as a semi-shade plant and radish as a sun plant, effects of growth light intensity on photosynthetic electron transport (PS) activity in chloroplasts and superoxide (O2.-) production in thylakoid membrane by irradiation were investigated. High-light chloroplasts of both plants showed higher PS activities than those grown under ow growth light intensity. High PS II and low PS I activities in ginseng chloroplasts (ratio of PS II/PS I : 1.1) were observed, but radish chloroplasts showed low PS II and high PS I activities (ratio of PS II/PS I : 0.3). PS II activity of both plants was little affected by temperature in range of 15-35$^{\circ}C$. Activities of whole -chain (PS II+I) in ginseng and PS I in radish were increased at high temperature (4$0^{\circ}C$). Preincubation of chloroplasts at 4$0^{\circ}C$ during 30 min, as a mild heat stress, caused rapid decrease in PS II and PS II+I activities of both plants. However PS I activity was not decreased in ginseng and rather increased in radish. O2.- production (NBT reduction) in Mehler reaction in the thylakoid membrane was inhibited by DCMU in both plants. DMBIB inhibited O2.- production in ginseng, but radish was insensitive to DMBIB. Electron flow system in ginseng thylakoid membrane was more susceptible to damage of photooxidation than that of radish.

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Stabilization of Membrane Proteins by Benzyladenine during Wheat Leaf Senescence (노쇠중인 밀잎에서 Benzyladenine에 의한 막단백질의 안정화)

  • 진창덕
    • Journal of Plant Biology
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    • v.35 no.2
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    • pp.117-123
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    • 1992
  • The effect of benzyladenine (BA) on lipid peroxidation and compositions of total insoluble proteins and chloroplast thylakoid protein from wheat primary leaves during senescence in the dark was studied. BA ($10^{-5}\;M$) treatment prevented conspicuously the loss of chlorophyll content and soluble and insoluble leaf protein contents in senescing wheat leaf segments during 4-day dark incubation. Under the BA treatment, especially, the level of insoluble protein was highly maintained than that of soluble protein. Also, the increase of malondialdehyde (MDA: the peroxidation product of membrane lipids) content was inhibited in the BA treated leaves. Three major polypeptide bands in quantity corresponding to 57, 26 and 12 KD molecular weight were clearly resolved with other minor bands by SDS-polyacrylamide gel electrophoresis (SDS-PAGE) in the insoluble protein fraction. The insoluble protein profiles of the control leaves showed a remarkable decrease in the intensity of the 57 and 12 KD band except for 26 KD band in the 72 h dark incubation. This loss during dark incubation was reduced by BA treatment. More than 20 polypeptides were resolved in the chloroplast thylakoid membrane fraction with the most prominent bands which are 59 and 57 KD ($\alpha\;and\;\beta$ subunit of coupling factor: CF) and 26 KD (apoprotein of LHCP). The changes in thylakoid protein profile during 72 h dark incubation showed the rapid degradation in control, but this degradation was prevented in quantity by BA treatment. The above results suggested that BA would inhibit the peroxidation of membrane lipids, thereby preventing the loss of membrane proteins which led to the maintenance of the membrane integrity including chloroplast thylakoid.

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LHCP phosphorylation and Chlorophyll-Fluorescence Quenching of PSII in Ginseng Thylakoid Members (인삼 틸라코이드에서 광계II의 LHCP 인산화와 형광 Quenching)

  • 양덕조;김명원
    • Journal of Ginseng Research
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    • v.16 no.2
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    • pp.124-128
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    • 1992
  • Vsing the phosphorylated thylakoid membrane induced by 5-35kLux of light intensities, we investigated the chlorophyll nuorescence quenching of PSII and the phosphorylation of LHCPII i in relation to the chlorophyll-bleaching of Panax ginseng C.A. Meyer. In the Presence of DCMU, both of the fluorescence yield of non-phosphorylated thylakoid and the rate of fluorescence quencing dependent on the Phosphorylation were high p. ginseng more then Glyine max L. And at the high light F intensity (above 25 kLux) the fluorescence quenching rate of p. ginseng compared with that of G. max reached nearly to 2 times. The LHCPII of P. ginseng was composed of 3 major Polypeptides (24.5, 26 and 27kD) and 3 minor polypeptides (24, 25.3 and 28.3kD) in the region of 24-29kD and differed, from G. max in both of the number and quantity of polypeptides. Among these polypeptides, the phosphorylated polypeptide dependent on the light intensity was 24kD in P. ginseng.

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Targeting Analysis of Lumenal Proteins of Chloroplast of Wheat using Proteomic Techniques

  • Kamal, Abu Hena Mostafa;Kim, Da-Eun;Oh, Myoung-Won;Chung, Keun-Yook;Cho, Yong-Gu;Kim, Hong-Sig;Song, Beom-Heon;Lee, Chul-Won;Uozumi, Nobuyuki;Choi, Jong-Soon;Cho, Kun;Woo, Sun-Hee
    • Proceedings of the Plant Resources Society of Korea Conference
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    • pp.14-14
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    • 2010
  • Plastid proteomics are essential organelles present in virtually all cells in plants and green algae. Plastids are responsible for the synthesis and storage of key molecules required for the basic architecture and functions of plant cells. The proteome of plastid, and in particular of chloroplast, have received significant amounts of attention in recent years. Various fractionation and mass spectrometry (MS) techniques have been applied to catalogue the chloroplast proteome and its sub-organelles compartments. To better understanding the function of the lumenal sub-organelles within the thylakoid network, we have carried out a systematical analysis and identification of the lumenal proteins in the thylakoid of wheat by using Tricine-SDS-PAGE, and LTQ-ESI-FTICR mass spectrometry followed by SWISS-PROT database searching. We isolation and fractionation these membrane from fully developed wheat leaves using a combination of differential and gradient centrifugation couple to high speed ultra-centrifuge. After collecting all proteins to eliminate possible same proteins, we estimated that there are 407 different proteins including chloroplast, chloroplast stroma, lumenal, and thylakoid membrane proteins excluding 20 proteins, which were identified in nucleus, cytoplasm and mitochondria. A combination of these three programs (PSORT, TargetP, TMHMM, and TOPPRED) was found to provide a useful tool for evaluating chloroplast localization, transit peptide, transmembranes, and also could reveal possible alternative processing sites and dual targeting. Finally, we report also sub-cellular location specific protein interaction network using Cytoscape software, which provides further insight into the biochemical pathways of photosynthesis. The present work helps understanding photosynthesis process in wheat at the molecular level and provides a new overview of the biochemical machinery of the thylakoid in wheat.

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The Isolation and Characterization of Chlorophyll-Protein Complexes in Thylakoid Membranes from Zoysia japonica and Spinach oleracea (잔디와 시금치의 Thylakoid Membrane으로부터 엽록소-단백질 복합체의 분리와 그 특성)

  • 김병규;장남기
    • Asian Journal of Turfgrass Science
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    • v.4 no.1
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    • pp.12-23
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    • 1990
  • The chlorophyll-protein complexes were separated from thylakoid membranes of Spinach oleracea and Zoysia japonica by two gel Systems of LiDodSO4-PAGE and LiDodSo4/Urea- PAGE under nondenaturing conditions. Seven chlorophyll~protein complexes of CPI*, CPI, CPII*. CP47, CP43, CP29 and CPII were fractionated from both S,oleracea and Zjaponica by LiDodSO4-PAGE. CPI, CP47 and CP43 contained more chlorophyll a than chlorophyll b. The patterns of their absorption spectra at room temperature were similliar to that of chlorophyll a, judging by their UV-spedtroscopy. On the other hand, CPII* and CPII contained approximately equim-olar quantities of chlorophyll a and b. Additional five chlorophyll-protein complexes not separated in the LiDodSO4-PAGE system were electrophoretically isolated from both S, oleracea and Zjaponica by LiDodSO4/Urca-PAGE. The chlorophyll-protein complex just above LRCII $\alpha$in the gel appears CCII-RC separeted recently. 23 kDa and 20 kDa cho-protein complexes is probably LHCIa and LHCIb as judged from their molecular weight. Two novel chlorophyll~protein complexes designated "CPI7" and "CPI6" were fractionate by this gel system. Their molecular weights respectively. Although the stoichiometry of their components and their roles in thylakoid membranes are not apparant, It is thought that they are another kinds of LHCI.other kinds of LHCI.

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Changes of Chloroplast Ultrastructure and Thylakoid Membrane Proteins during Growth of Ginseng (Panax ginseng C.A. Meyer) Leaf (인삼(Panax ginseng C.A. Meyer) 잎의 생장과정에 따른 엽록체 미세구조 및 틸라코이드막 단백질의 변화)

  • Ahn, Joung-Sook;Park, Hoon;Kim, Woo-Kap
    • Journal of Ginseng Research
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    • v.19 no.3
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    • pp.275-280
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    • 1995
  • The formation of thylakoid membrane proteins and changes in the chloroplast ultrastructure of ginseng leaf were investigated as a function of time following the leaf emergence. The leaf chloroplast obtained just after the leaf emergence showed short rod-like thylakoids which were connected and arranged in 3~4 layers along the longitudinal axis of the chloroplast. The 10 DAE (days after emergence) chloroplast started to form grana structure. The typical grana structure was observed 17 DAE, and the grana was fully developed 28 DAE. The membrane proteins obtained from just after emerging leaf were separated into many minor bands indicating no CP-complex formation yet. LHC II was detected after 10 days. CP 47 and CP 43 were detected after 17 days. After 28 days, the PS I and PS II proteins were distinctly separated into CP 1, LHC II, CP 47, CP 43, CP 29, CP 27+24. Thus, the appearance of the light harvesting protein, LHC II, which was concentrated in grana stacks, was consis tent in time with the formation of grana stacks 17 DAE. Key words Chloroplast ultrastructure, grana, CP-complex, LHC II.

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Effects of Spermine on Changes in Chlorophyll-Protein Complexes and Plastic Membrane Proteins of Mung Bean Cotyledons during Greening (녹화중인 녹두 자엽의 엽록소-단백질 복합체 및 색소체막 단백질의 변화에 미치는 Spermine의 효과)

  • 홍정희;박흥덕
    • Journal of Environmental Science International
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    • v.4 no.4
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    • pp.335-344
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    • 1995
  • Developmental changes of chlorophyll-protein complexes (CPs) and plastid membrane proteins in greening mung bean cotyledons and the effect of spermine therein were examined by SDS-polyacrylamide gel electrophoresis. The changes in the amounts of CPs became larger with the progress of greening and light-harvesting chlorophyll a/b protein (LHCP) was the main CP in the early greening stage up to f h. As the greening proceeded, chlorophyll-protein of the photosystem I (CPI) accumulated. Application of spermine were effective in accumulating CPs of the thylakoid membrane in the early phase of greening. In the profiles of the plastid membrane proteins, quantitative and qualitative changes were observed with the onset of greening up to 72 h. 56 kD protein of major intensity was observed in all greened chloroplasts and 24 kD protein increased remarkablly in both control and spermine-treated cotyledons. The thylakoids from spermine-treated cotyledons showed hither amounts of thylakoid proteins as compared to the controls. The results suggest that spermine may play a role in the regulation of plastid development and stabilizes the membrane function during greening.

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Characteristics of Light Harvesting Chlorophyll-Protein Complex and Singlet Oxygen ($^1O_2$) Quenching in Leaf-burning Disease from Panax ginseng C. A. Meyer (인삼 Light Harvesting Chlorophyll Protein의 특성 및 엽소병에서 Singlet Oxygen($^1O_2$) Quenching)

  • 양덕조;이성택
    • Journal of Ginseng Research
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    • v.13 no.2
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    • pp.158-164
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    • 1989
  • In order to determine the relationships between the lea(-burning disease and the light harvesting chlorophyll-protein (LHCP) complex in Panax ginseng C. A. Meyer, we investigated the chlorophyll-protein (CP) complex of the thylakoid membrane and its characteristics. In P. ginseng four Cp-complex bands determined by non-denaturing SDS-PAGE were identified CP I'(containing reaction center of photosystem I and LHCP I antennae), CP I (reaction center of photosystem I) LHCP II** (oligoform of LHCP II), and LHCP II (photosystem II antennae, CP 26 and CP 29) by Bassis and Dunahay's procedures. Under our experimental condition, the CP I band was only observed in P. ginseng and the band intensity of LHCP II** in P ginseng was higher than in spinach and soybean. There were differences in the absorption and fluorescence spectra and chlorophyll a/b ratio of the CP-complex bands between P. ginseng and other Plants. The Polypeptidr content of P. ginseng thylakoid was lower than in spinach and soybean thylakoid, and the Polypeptide profiles of P. ginseng was low band intensity, especially about 29-35 kD, 55 kD, and 60 kD, compared to spinach and soybean. The inhibitory effects of 2,5-dimethylfuran, specific singlet oxygen ($^1O_2$) quencher, showed that singlet oxygen destroyed 60% of chl.a, 90% of chl.b and 70% of carotenoid in bleaching P. ginseng with leaf-burning disease.

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