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A heterogeneous poly A-mRNA fraction was isolated from rat liver microsomes by phenol:chloroform extraction, millipore filtration, and poly U-agarose affinity chromatography. The fractions were characterized by their secondary structures and poly A contents. From translational studies, the isolated fraction was found to have high glutamate dehydrogenase template activity in cell-free systems containing microsomes or polysomes. A spectrophotometric procedure for following enzyme biosynthesis was also developed. 相似文献
4.
The catalase-peroxidase hydroperoxidase I of Escherichia coli has been confirmed to be located in the cytoplasm using two independent methods. Catalase activity was found predominantly (> 95%) in the cytoplasmic fraction following spheroplast formation. The cytoplasmic enzyme glucose-6-phosphate dehydrogenase and the periplasmic enzyme alkaline phosphatase were used as controls. The second method of immunogold staining for the enzyme in situ revealed an even distribution of the enzyme across the cell. 相似文献
5.
Simoyi MF Falkenstein E Van Dyke K Blemings KP Klandorf H 《Comparative biochemistry and physiology. Part B, Biochemistry & molecular biology》2003,135(2):325-335
Urate oxidase is not present in birds yet allantoin, a product of this enzyme, has been measured in birds. Studies were designed to compare the concentrations of plasma purine derivatives in chickens and turkeys fed inosine-supplemented diets. The first study consisted of 12 male chicks that were fed diets supplemented with 0.6 mol inosine or hypoxanthine per kilogram diet from 3- to 6-week-old. Study 2 consisted of 12 turkey poults (toms) fed inosine-supplemented diets (0.7 mol/kg) from 6- to 8-week-old. Plasma allantoin and oxypurines concentrations were measured weekly using high performance liquid chromatography. Plasma uric acid (PUA) in chickens fed inosine-supplemented diets increased from 0.31 to 1.34 mM (P<0.05) at the end of week 2. In turkeys, those fed control diet had 0.17 mM PUA concentration compared to 0.3 mM in those fed the inosine diet at week 2 (P<0.05). Allantoin concentration increased in chickens from week 1 to 2 while a decrease was observed in turkeys (P<0.005) for both treatments. These data show that allantoin is present in turkey and chicken plasma. The presence of allantoin in avian plasma is consistent with uric acid acting as an antioxidant in these species. 相似文献
6.
Oscillations in photosynthesis are initiated and supported by imbalances in the supply of ATP and NADPH to the Calvin cycle 总被引:4,自引:0,他引:4
Agu Laisk Katharina Siebke Ulvi Gerst Hillar Eichelmann Vello Oja Ulrich Heber 《Planta》1991,185(4):554-562
Oscillations in the rate of photosynthesis of sunflower (Helianthus annuus L.) leaves were induced by subjecting leaves, whose photosynthetic apparatus had been activated, to a sudden transition from darkness or low light to high-intensity illumination, or by transfering them in the light from air to an atmosphere containing saturating CO2. It was found that at the first maximum, light-and CO2-saturated photosynthesis can be much faster than steady-state photosynthesis. Both QA in the reaction center of PS II and P700 in the reaction center of PS I of the chloroplast electron-transport chain were more oxidized during the maxima of photosynthesis than during the minima. Maxima of P700 oxidation slightly preceded maxima in photosynthesis. During a transition from low to high irradiance, the assimilatory force FA, which was calculated from ratios of dihydroxyacetone phosphate to phosphoglycerate under the assumption that the reactions catalyzed by NADP-dependent glyceraldehydephosphate dehydrogenase, phosphoglycerate kinase and triosephosphate isomerase are close to equilibrium, oscillated in parallel with photosynthesis. However, only one of its components, the calculated phosphorylation potential (ATP)/(ADP)(Pi), paralleled photosynthesis, whereas calculated NADPH/NADP ratios exhibited antiparallel behaviour. When photosynthetic oscillations were initiated by a transition from low to high CO2, the assimilatory force FA declined, was very low at the first minimum of photosynthesis and increased as photosynthesis rose to its second maximum. The observations indicate that the minima in photosynthesis are caused by lack of ATP. This leads to overreduction of the electron-transport chain which is indicated by the reduction of P700. During photosynthetic oscillations the chloroplast thylakoid system is unable to adjust the supply of ATP and NADPH rapidly to demand at the stoichiometric relationship required by the carbonreduction cycle.Abbreviations PGA
3-phosphoglycerate
- DHAP
dihydroxyacetone phosphate
- P700
electron-donor pigment in the reaction enter of PS I
- QA
quinone acceptor in the reaction center of PS II
This work received support from the Estonian Academy of Sciences, the Bavarian Ministry of Science and Art and the Sonderforschungsbereich 251 of the University of Würzburg. We are grateful for criticism by D.A. Walker, Robert Hill Institute, University of Sheffield, U.K. and by Mark Stitt, Institute of Botany, University of Heidelberg, FRG. 相似文献
7.
Vello Oja Hillar Eichelmann Agu Anijalg Heikko Rämma Agu Laisk 《Photosynthesis research》2010,103(3):153-166
Oxidation of photosystem I (PSI) donors under far-red light (FRL), slow re-reduction by stromal reductants and fast re-reduction
in the dark subsequent to illumination by white light (WL) were recorded in leaves of several C3 plants at 810 and 950 nm. During the re-reduction from stromal reductants the mutual interdependence of the two signals followed
the theoretical relationship calculated assuming redox equilibrium between plastocyanin (PC) and P700, with the equilibrium
constant of 40 ± 10 (ΔE
m = 86–99 mV) in most of the measured 24 leaves of nine plant species. The presence of non-oxidizable PC of up to 13% of the
whole pool, indicating partial control of electron transport by PC diffusion, was transiently detected during a saturation
pulse of white light superimposed on FRL or on low WL. Nevertheless, non-oxidizable PC was absent in the steady state during
fast light-saturated photosynthesis. It is concluded that in leaves during steady state photosynthesis the electron transport
rate is not critically limited by PC diffusion, but the high-potential electron carriers PC and P700 remain close to the redox
equilibrium. 相似文献
8.
Membrane transporter ProP from Escherichia coli senses extracellular osmolality and responds by mediating the uptake of osmoprotectants such as glycine betaine when osmolality is high. Earlier EPR and NMR studies showed that a peptide replica of the cytoplasmic ProP carboxyl terminus (residues D468-R497) forms a homodimeric, antiparallel, alpha-helical coiled coil in vitro stabilized by electrostatic interactions involving R488. Amino acid replacement R488I disrupted coiled-coil formation by the ProP peptide, elevated the osmolality at which ProP became active, and rendered the osmolality response of ProP transient. In the present study, either E480 or K473 was replaced with cysteine (Cys) in ProP, a Cys-less, fully functional, histidine-tagged ProP variant, to use Cys-specific cross-linking approaches to determine if antiparallel coiled-coil formation and dimerization of the intact protein occur in vivo. The Cys at positions 480 would be closer in an antiparallel dimer than those at positions 473. These replacements did not disrupt coiled-coil formation by the ProP peptide. Partial homodimerization of variant ProP-E480C could be demonstrated in vivo and in membrane preparations via Cys-specific cross-linking with dithiobis(maleimidoethane) or by Cys oxidation to cystine by copper phenanthroline. In contrast, these reagents did not cross-link ProP with Cys at position 133 or 241. Cross-linking of ProP with Cys at position 473 was limited and occurred only if ProP was overexpressed, consistent with an antiparallel orientation of the coiled coil in the intact protein in vivo. Although replacement E480C did not alter transporter activity, replacement K473C reduced the extent and elevated the threshold for osmotic activation. K473 may play a role in ProP structure and function that is not reflected in altered coiled-coil formation by the corresponding peptide. Substitution R488I affected the activities of ProP-(His)(6), ProP-E480C, and ProP-K473C as it affected the activity of ProP. Surprisingly, it did not eliminate cross-linking of Cys at position 480, and it elevated cross-linking at position 473, even when ProP was expressed at physiological levels. This suggested that the R488I substitution may have changed the relative orientation of the C-termini within the dimeric protein from antiparallel to parallel, resulting in only transient osmotic activation. These results suggest that ProP is in monomer-dimer equilibrium in vivo. Dimerization may be mediated by C-terminal coiled-coil formation and/or by interactions between other structural domains, which in turn facilitate C-terminal coiled-coil formation. Antiparallel coiled-coil formation is required for activation of ProP at low osmolality. 相似文献
9.
A computer-operated routine of gas exchange and optical measurements to diagnose photosynthetic apparatus in leaves 总被引:10,自引:4,他引:6
A.Laisk V.Oja B.Rasulov H.Rämma H.Eichelmann I.Kasparova H.Pettai E.Padu & E.Vapaavuori 《Plant, cell & environment》2002,25(7):923-943
Photosynthesis is a complex process whose rate is affected by many biochemical and biophysical factors. Fortunately, it is possible to determine, or at least estimate, many of the most important parameters using a combination of optical methods and gas transient analyses. We describe here a computer‐operated routine that has been developed to make detailed assessments of photosynthesis at a comprehensive level. The routine comprised the following measurements: steady‐state light and CO2 response curves of net CO2 assimilation at 21 and 2 kPa O2; transients from limiting to different saturating CO2 concentrations at 2 kPa O2; post‐illumination CO2 fixation transient; dark–light induction of O2 evolution; O2 yield from one saturating single‐turnover flash; chlorophyll fluorescence F0, Fs and Fm during the light and CO2 response curves; leaf transmission at 820 nm (P700+) during the light and CO2 response curves; post‐illumination re‐reduction time of P700+. The routine was executed on a two‐channel fast‐response gas exchange measurement system (A. Laisk and V. Oja: Dynamic Gas Exchange of Leaf Photosynthesis. CSIRO, Canberra, Australia). Thirty‐six intrinsic characteristics of the photosynthetic machinery were derived, including quantum yield of CO2 fixation (YCO2), time constant of P700 re‐reduction (τ′), relative optical cross‐sections of PSII and PSI antennae (aII, aI), PSII and PSI density per leaf area unit, plastoquinone pool, total mesophyll resistance, mesophyll diffusion resistance, Vm, Km(CO2) and CO2/O2 specificity of Rubisco, RuBP pool at CO2 limitation (assimilatory charge). An example of the routine and calculations are shown for one leaf and data are presented for leaves of 8‐year‐old‐trees of two birch clones growing in Suonenjoki Forest Research Station, Finland, during summer 2000. Parameters YCO2, basic τ′, aII, aI, Km(CO2) and Ks varied little in different leaves [relative standard deviation (RSD) < 7%], other parameters scattered widely (RSD typically 10–40%). It is concluded that the little scattered parameters are determined by basic physico‐chemical properties of the photosynthetic machinery whereas the widely scattered parameters are adjusting to growth conditions. The proposed non‐destructive routine is suitable for diagnosing the photosynthetic machinery of leaves and may be applied in plant ecophysiology and in genetic engineering of plants. 相似文献
10.
M Hillar 《Archives internationales de physiologie et de biochimie》1978,86(2):227-233
Histone blocks proton uptake by mitochondria incubated in the presence of valinomycin or DNP. In the presence of DNP valinomycin-induced H+ uptake is not affected by histone. H+ uptake induced by nigericin is not affected by histone as well. Postulated mechanism of histone action involves the immobilization of proton translocation in mitochondrial membrane and induction of local change in H+ concentration, the prevention of the interaction between H+ and natural K+-carrier and Mg2+ transport system or valinomycin. 相似文献