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1.
Elanskaya IV Grivennikova VG Groshev VV Kuznetsova GV Semina ME Timofeev KN 《Biochemistry. Biokhimii?a》2004,69(2):137-142
Insertion mutant Ins2 of the cyanobacterium Synechocystis sp. PCC 6803, lacking NAD(P)H:quinone oxidoreductase (NQR) encoded by drgA gene, was characterized by higher sensitivity to quinone-type inhibitors (menadione and plumbagin) than wild type (WT) cells. In photoautotrophically grown cyanobacterial cells more than 60% of NADPH:quinone-reductase activity, as well as all NADPH:dinoseb-reductase activity, was associated with the function of NQR. NQR activity was observed only in soluble fraction of cyanobacterial cells, but not in membrane fraction. The effects of menadione and menadiol on the reduction of Photosystem I reaction center (P700(+)) after its photooxidation in the presence of DCMU were studied using the EPR spectroscopy. The addition of menadione increased the rate of P700(+) reduction in WT cells, whereas in Ins2 mutant the reduction of P700(+) was strongly inhibited. In the presence of menadiol the reduction of P700(+) was accelerated both in WT and Ins2 mutant cells. These data suggest that NQR protects the cyanobacterial cells from the toxic effect of exogenous quinones by their reduction to hydroquinones. These data may also indicate the probable functional homology of Synechocystis sp. PCC 6803 NQR with mammalian and plant NAD(P)H:quinone oxidoreductases (DT-diaphorases). 相似文献
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《Bioscience, biotechnology, and biochemistry》2013,77(12):3180-3188
Cyclic electron transport and NADH and/or NADPH (NAD(P)H)-oxidizing activities were investigated in Synechocystis sp. PCC6803 grown under various stressed conditions and in ndhB-less (M55) and ycf33-deletion mutants. Activity staining and inhibitor data suggested that the ferredoxin-quinone reductase (FQR) route is the main pathway in ycf33-deletion and high-light (300 μE m?2 s?1)-grown cells as well as in M55 cells. The FQR route was highly sensitive to HgCl2, but not to diphenyleneiodonium (DPI). On the other hand, cells grown under low CO2 (0.03%) or normal (100 μE m?2 s?1, 3% CO2) conditions were found perhaps to use the complex I-type NAD(P)H dehydrogenase route, which was found to be highly sensitive to DPI but not to HgCl2. In high-salt (0.55 M NaCl)-grown cells, the amount of ferredoxin-NADP+ oxidoreductase (FNR) increased, and the main cyclic electron flow was perhaps the FNR route. Both DPI and HgCl2 were strong inhibitors of the FNR route. 相似文献
4.
Kinetics of the redox reactions in the reaction center (P700) of photosystem I (PSI) of the cyanobacterium Synechocystis sp. PCC 6803 have been studied by EPR spectroscopy. The redox kinetics were recorded based on accumulation of the EPRI signal when the final signal was the sum of individual signals produced in response to illumination of the cells. After prolonged (more than 3 sec) dark intervals between illuminations, the kinetic curve of the EPR signal from P700+ was multiphasic. After a sharp increase in the signal amplitude at the beginning of illumination (phase I), the amplitude rapidly (for 0.1-0.2 sec) decreased (phase II). Then the signal amplitude gradually increased (phase III) until the steady rate of electron transfer was established. With short-term (1 sec) dark intervals between the flashes and also in the presence of 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU), the kinetics of the light-induced increase in the EPR signal from P700+ were monophasic. Inhibition with iodoacetamide of electron transport on the acceptor side of PSI under anaerobic conditions or an increase in the amount of respiration substrates on addition of glucose into a suspension of DCMU-treated wild-type cells increased the level of P700 reduction in phase III. The findings suggest that the kinetic curve of the EPR signal from P700+ is determined by both the electron entrance onto P700+ on the donor side of PSI and activity of electron acceptors of PSI. 相似文献
5.
A recent proteomic analysis of the thylakoid lumen of Arabidopsis thaliana revealed the presence of several PsbP-like proteins, and a homologue to this gene family was detected in the genome of the cyanobacterium Synechocystis sp. PCC 6803 (Schubert M, Petersson UA, Haas BJ, Funk C, Schröder WP, Kieselbach T (2002) J Biol Chem 277, 8354–8365). Using a peptide-directed antibody against this cyanobacterial PsbP-like protein (sll1418) we could show that it was localized in the thylakoid membrane and associated with Photosystem II. While salt washes did not remove the PsbP-like protein from the thylakoid membrane, it was partially lost during the detergent-based isolation of PSII membrane fractions. In total cell extracts this protein is present in the same amount as the extrinsic PsbO protein. We did not see any significant functional difference between the wild-type and a PsbP-like insertion mutant. 相似文献
6.
利用离子交换与凝胶过滤层析 ,从n dodecylβ D maltoside(DM)处理的集胞蓝藻SynechocystisPCC6 80 3细胞粗提液中 ,首次分离到两个包含NDH疏水亚基NdhA的亚复合体。酶活性分析表明 ,分离到的NDH亚复合体具有NADPH 氮蓝四唑 (NBT)氧化还原酶活性 ,以NADPH为电子供体可以还原铁氰化钾、二溴百里香醌 (DBMIB)、二氯酚靛酚 (DCPIP)、duroquinone以及UQ 0等质醌类电子受体。 相似文献
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The PsbP-like protein of the cyanobacterium Synechocystis sp. PCC 6803 is a peripheral component of Photosystem II, located at the lumenal side of the thylakoid membrane. Removal of this protein leads to decreased competitive potential of a PsbP-like deletion mutant when grown in a mixture with wild-type cells. Flash-induced oxygen evolution traces of the mutant show a higher probability of misses, correlated with increased amplitudes of the S-states decay in the dark. Thermoluminescence emission traces demonstrate a changed charge recombination pattern in the mutant, the S(3)Q(B)(-) couple becoming the major species instead of the S(2)Q(B)(-). Our data suggest a possible role of the PsbP-like protein in stabilisation of the charge separation in Photosystem II of cyanobacteria through interaction with the Mn cluster. 相似文献
8.
Naoyuki Tajima Shusei Sato Fumito Maruyama Takakazu Kaneko Naobumi V. Sasaki Ken Kurokawa Hiroyuki Ohta Yu Kanesaki Hirofumi Yoshikawa Satoshi Tabata Masahiko Ikeuchi Naoki Sato 《DNA research》2011,18(5):393-399
Synechocystis sp. PCC 6803 is the most popular cyanobacterial strain, serving as a standard in the research fields of photosynthesis, stress response, metabolism and so on. A glucose-tolerant (GT) derivative of this strain was used for genome sequencing at Kazusa DNA Research Institute in 1996, which established a hallmark in the study of cyanobacteria. However, apparent differences in sequences deviating from the database have been noticed among different strain stocks. For this reason, we analysed the genomic sequence of another GT strain (GT-S) by 454 and partial Sanger sequencing. We found 22 putative single nucleotide polymorphisms (SNPs) in comparison to the published sequence of the Kazusa strain. However, Sanger sequencing of 36 direct PCR products of the Kazusa strains stored in small aliquots resulted in their identity with the GT-S sequence at 21 of the 22 sites, excluding the possibility of their being SNPs. In addition, we were able to combine five split open reading frames present in the database sequence, and to remove the C-terminus of an ORF. Aside from these, two of the Insertion Sequence elements were not present in the GT-S strain. We have thus become able to provide an accurate genomic sequence of Synechocystis sp. PCC 6803 for future studies on this important cyanobacterial strain. 相似文献
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An NADPH-specific NDH-1 sub-complex was separated by native-polyacrylamide gel electrophoresis and detected by activity staining from the whole cell extracts of Synechocystis PCC6803. Low CO2 caused an increase in the activity of this sub-complex quickly, accompanied by an evident increase in the expression of NdhK and PSI-driven NADPH oxidation activity that can reflect the activity of NDH-1-mediated cyclic electron transport. During incubation with high CO2, the activities of NDH-1 sub-complex and PSI-driven NADPH oxidation as well as the protein level of NdhK slightly increased at the beginning, but decreased evidently in various degrees along with incubation time. These results suggest that CO2 concentration in vitro as a signal can control the activity of NDH-1 complex, and NDH-1 complex may in turn function in the regulation of CO2 uptake. 相似文献
10.
Robert Jeanjean Sylvie Bédu Michel Havaux Hans C.P Matthijs Françoise Joset 《FEMS microbiology letters》1998,167(2):131-137
The cyanobacterium Synechocystis PCC6803 induces a photosystem I cyclic electron transfer route independent of type 1 NAD(P)H dehydrogenase. The capacity to tolerate raised salinity conditions was shown to operate in a mutant lacking functional type 1 NAD(P)H dehydrogenase. The mutant showed salt-induced enhancement of photosystem I cyclic electron transfer and respiratory capacities. Moreover, this salt-adapted energetic state also restored the capacity of the mutant to grow under inorganic carbon limitation. Uptake of the latter in these conditions became almost as efficient as in the wild-type. The acquired energetic capacities, in contrast, did not allow restoration of photoheterotrophic growth in the type 1 NAD(P)H dehydrogenase mutant. 相似文献
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Kufryk G Hernandez-Prieto MA Kieselbach T Miranda H Vermaas W Funk C 《Photosynthesis research》2008,95(2-3):135-145
The cyanobacterial small CAB-like proteins (SCPs) are one-helix proteins with compelling similarity to the first and third
transmembrane helix of proteins belonging to the CAB family of light-harvesting complex proteins in plants. The SCP proteins
are transiently expressed at high light intensity and other stress conditions but their exact function remains largely unknown.
Recently we showed association of ScpD with light-stressed, monomeric Photosystem II in Synechocystis sp. PCC 6803 (Yao et al. J Biol Chem 282:267–276, 2007). Here we show that ScpB associates with Photosystem II at normal
growth conditions. Moreover, upon introduction of a construct into Synechocystis so that ScpB is expressed continuously under normal growth conditions, ScpE was detected under non-stressed conditions as
well, and was copurified with tagged ScpB and Photosystem II. We also report on a one-helix protein, Slr1544, that is somewhat
similar to the SCPs and whose gene is cotranscribed with that of ScpD; Slr1544 is another member of the extended light-harvesting-like
(Lil) protein family, and we propose to name it LilA.
Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.
Galyna Kufryk and Miguel Hernandez-Prieto have contributed equally to this work. 相似文献
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藻胆体是蓝藻细胞主要的捕光天线色素超分子复合体,主要由核心体和外围的杆两部分组成,核心体主要由别藻蓝蛋白组装而成,参与光能向光合作用反应中心的传递.该研究通过PCR扩增出集胞藻6803别藻蓝蛋白α亚基(ApcA)编码基因apcA,构建表达质粒pET-32a(+)-apcA,并将其转入大肠杆菌BL21(DE3)pLysS菌株中;通过IPTG诱导表达重组蛋白,并利用组氨酸标签将可溶性目的蛋白进行亲和纯化后,免疫日本大耳白兔,从而获得多克隆抗体.间接ELISA法揭示ApcA抗体效价可高达1∶1 025 000;蛋白免疫印迹确定该抗体具有高度特异性.表明该研究成功制备了集胞藻6803藻胆体别藻蓝蛋白多克隆抗体,为进一步研究藻胆体的核心体在光能传递过程中所承担的重要生理角色奠定了生化基础. 相似文献
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PCR扩增了蓝细菌集胞藻6803(Synechocystis sp.PCC6803)的agp基因(编码ADP-葡萄糖焦磷酸羧化酶),进一步以pUC118为载体将其克隆到大肠杆菌中,构建了pUCA质粒。通过DNA体外重组,以红霉素抗性基因部分取代agp基因片段,构建了既含agp基因上游及下游序列、又携带选择性标记-红霉素抗性的pUCAE质粒。该质粒转化野生型集胞藻6803细胞,获得了能在含红霉素的培养基上正常生长的agp基因缺失突变株。对该突变株基因组DNA进行PCR扩增,验邝了其基因结构的正确性。突变株细胞生长速度较野生型细胞快,胞内的叶绿素含量比野生型细胞高,表明该突变株具有较高的光合效率。在突变株中未检测到糖原的存在,进一步从生理水平上验证了突变株构建的正确性。 相似文献
16.
Bishop CL Ulas S Baena-Gonzalez E Aro EM Purton S Nugent JH Mäenpää P 《Photosynthesis research》2007,93(1-3):139-147
The psbZ gene of Synechocystis sp. PCC 6803 encodes the ∼6.6 kDa photosystem II (PSII) subunit. We here report biophysical, biochemical and in vivo characterization
of Synechocystis sp. PCC 6803 mutants lacking psbZ. We show that these mutants are able to perform wild-type levels of light-harvesting, energy transfer, PSII oxygen evolution,
state transitions and non-photochemical quenching (NPQ) under standard growth conditions. The mutants grow photoautotrophically;
however, their growth rate is clearly retarded under low-light conditions and they are not capable of photomixotrophic growth.
Further differences exist in the electron transfer properties between the mutants and wild type. In the absence of PsbZ, electron
flow potentially increased through photosystem I (PSI) without a change in the maximum electron transfer capacity of PSII.
Further, rereduction of P700+ is much faster, suggesting faster cyclic electron flow around PSI. This implies a role for PsbZ in the regulation of electron
transfer, with implication for photoprotection. 相似文献
17.
The role of putative Na+/H+ antiporters encoded by nhaS1 (slr1727), nhaS3 (sll0689), nhaS4 (slr1595), and nhaS5 (slr0415) in salt stress response and internal pH regulation of the cyanobacterium Synechocystis PCC 6803 was investigated. For this purpose the mutants (single, double, and triple) impaired in genes coding for Na+/H+ antiporters were constructed using the method of interposon mutagenesis. PCR analyses of DNA demonstrated that mutations in nhaS1, nhaS4, and nhaS5 genes were segregated completely and the mutants contained only inactivated copies of the corresponding genes. Na+/H+ antiporter encoded by nhaS3 was essential for viability of Synechocystis since no completely segregated mutants were obtained. The steady-state intracellular sodium concentration and Na+/H+ antiporter activities were found to be the same in the wild type and all mutants. No differences were found in the growth rates of wild type and mutants during their cultivation in liquid media supplemented with 0.68 M or 0.85 M NaCl as well as in media buffered at pH 7.0, 8.0, or 9.0. The expression of genes coding for Na+/H+ antiporters was studied. No induction of any Na+/H+ antiporter encoding gene expression was found in wild type or single mutant cells grown under high salt or at different pH values. Nevertheless, in cells of double and triple mutants adapted to high salt or alkaline pH some of the remaining Na+/H+ antiporter encoding genes showed induction. These results might indicate that some of Na+/H+ antiporters can functionally replace each other under stress conditions in Synechocystis cells lacking the activity of more than one antiporter. 相似文献
18.
Danika Trautmann Bj?rn Vo? Annegret Wilde Salim Al-Babili Wolfgang R. Hess 《DNA research》2012,19(6):435-448
Synechocystis sp. PCC 6803 is a widely used model cyanobacterium for studying photosynthesis, phototaxis, the production of biofuels and many other aspects. Here we present a re-sequencing study of the genome and seven plasmids of one of the most widely used Synechocystis sp. PCC 6803 substrains, the glucose tolerant and motile Moscow or ‘PCC-M’ strain, revealing considerable evidence for recent microevolution. Seven single nucleotide polymorphisms (SNPs) specifically shared between ‘PCC-M’ and the ‘PCC-N and PCC-P’ substrains indicate that ‘PCC-M’ belongs to the ‘PCC’ group of motile strains. The identified indels and SNPs in ‘PCC-M’ are likely to affect glucose tolerance, motility, phage resistance, certain stress responses as well as functions in the primary metabolism, potentially relevant for the synthesis of alkanes. Three SNPs in intergenic regions could affect the promoter activities of two protein-coding genes and one cis-antisense RNA. Two deletions in ‘PCC-M’ affect parts of clustered regularly interspaced short palindrome repeats-associated spacer-repeat regions on plasmid pSYSA, in one case by an unusual recombination between spacer sequences. 相似文献
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Sandra Turconi Jochen Kruip Gerd Schweitzer Matthias Rögner Alfred R. Holzwarth 《Photosynthesis research》1996,49(3):263-268
Picosecond time-resolved fluorescence measurements have been performed as a function of emission wavelengths in order to investigate the possible functional differences between monomeric and trimeric Photosystem I (PS I) particles from a cyanobacterium Synechocystis. Applying global analysis, four kinetic components were found necessary to describe the fluorescecne decay for both monomers and trimers of PS I. The lifetimes and spectra of the respective components are quite similar, indicating that they can be attributed to identical processes in both the monomers and trimers. It is concluded that both forms of PS I are capable of efficient energy transfer and charge separation, in agreement with a physiological role of both forms. Small differences in the fluorescence decays are discussed in terms of a slightly higher ratio of red emitting pigments per reaction centre in trimers of PS I. A comparison to Synechococcus PS I particles reveals the higher red chlorophyll content of the latter.Abbreviations -DM-
-dodecyl-maltoside
- Chl-
chlorophyll
- CMC-
critical micellar concentration
- DAS-
decay-associated spectrum
- DCM-
4-dicyano-methylene-2-methyl-6-(-dimethyl-aminostyryl)-4h-pyran
- FWHM-
full-width at half-maximum
- P700-
primary electron donor of Photosystem I
- PS-
photosystem
- RC-
reaction centre 相似文献
20.
小鼠金属硫蛋白在聚胞藻中的金属诱导表达与纯化 总被引:12,自引:0,他引:12
应用蓝藻类金属硫蛋白基因启动子(smt O-P)的金属诱导性,在单细胞的聚胞藻PCC 6803中表达小鼠金属硫蛋白结构基因(mMT-1 cDNA)。在大肠杆菌HB 101中构建含有smt O-P和mMT1 cDNA的穿梭表达载体pKT-MRE,经质粒转移,链霉素筛选,Southern和Western杂交分析鉴定得稳定的转基因工程藻落。同时,做小批量锌诱导表达,并纯化了外源蛋白,5L培养液含鲜藻重5.0g,得到3.5mg mMT-1;转基因藻在高金属浓度下的耐受性测定表明,外源基因的表达提高了蓝藻对金属离子的抗性,约为野生藻的2倍。 相似文献