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161.
The cellular pathway of sugar uptake in developing cotyledons of Vicia faba L. and Phaseolus vulgaris L. seed was evaluated using a physiological approach. The cotyledon interface with the seed coat is characterised by a specialised dermal cell complex. In the case of Vicia faba cotyledons, the epidermal component of the dermal cell complex is composed of transfer cells. Sucrose is the major sugar presented to the outer surface of both cotyledons and it is taken up from the apoplasm unaltered. Estimated sucrose concentrations within the apparent free space of Vicia and Phaseolus cotyledons were 105 and 113 mM respectively. Rates of in-vitro uptake of [14C]sucrose by cotyledon segments or by whole cotyledons following physical removal or porter inactivation of the outer cells demonstrated that, for both Vicia and Phaseolus cotyledons, the dermal cell complexes are the most intense sites of sucrose uptake. Accumulation of [14C]sucrose in the storage parenchyma of whole cotyledons was directly affected by experimental manipulation of uptake by the outer cell layers and plasmolytic disruption of the interconnecting plasmodesmata. These findings indicated that sucrose accumulated by the dermal cell complexes is transported symplasmically to the storage parenchyma. Overall, it is concluded that the dermal cell complexes of the developing legume embryo, irrespective of the presence or absence of wall ingrowths, are the major sites for the uptake of sucrose released from the maternal tissues to the seed apoplasm. Thereafter, the accumulated sucrose is transported radially inward through the symplast to the storage parenchyma.Abbreviations AFS
apparent free space
- CF
5-(6)-carboxyfluorescein
- CFDA
5-(6)-carboxyfluorescein diacetate
- Mes
2-(N-morpholino)ethanesulfonic acid
- PCMBS
p-chloromercuribenzenesulfonic acid
- SRG
sulphorhodamine G
The investigation was supported by funds from the Research Management Committee, The University of Newcastle and the Australian Research Council. One of us, R. McDonald, gratefully acknowledges the support of an Australian Postgraduate Research Award. We are grateful to Stella Savoury for preparing the photomicrographs. 相似文献
162.
T. Bei-Paraskevopoulou R. Anastassiou J. Argyroudi-Akoyunoglou 《Photosynthesis research》1995,44(1-2):93-106
The appearance of the light harvesting II (LHC II) protein in etiolated bean leaves, as monitored by immunodetection in LDS-solubilized leaf protein extracts, is under phytochrome control. A single red light pulse induces accumulation of the protein, in leaves kept in the dark thereafter, which follows circadian oscillations similar to those earlier found for Lhcb mRNA (Tavladoraki et al. (1989) Plant Physiol 90: 665–672). These oscillations are closely followed by oscillations in the capacity of the leaf to form Chlorophyll (Chl) in the light, suggesting that the synthesis of the LHC II protein and its chromophore are in close coordination. Experiments with levulinic acid showed that PChl(ide) resynthesis does not affect the LHC II level nor its oscillations, but new Chl a synthesis affects LHC II stabilization in thylakoids, implicating a proteolytic mechanism. A proteolytic activity against exogenously added LHC II was detected in thylakoids of etiolated bean leaves, which was enhanced by the light pulse. The activity, also under phytochrome control, was found to follow circadian oscillations in verse to those in the stabilization of LHC II protein in thylakoids. Such a proteolytic mechanism therefore, may account for the circadian changes observed in LHC II protein level, being implicated in pigment-protein complex assembly/stabilization during thylakoid biogenesis.Abbreviations Chl
chlorophyll
- CL
continuous light
- D
dark
- FR
far-red light
- LA
levulinic acid
- LHC II
light-harvesting complex serving Photosystem II
- PChl(ide)
protochlorophyllide
- PCR
protochlorophyllide oxidoreductase
- R
red light 相似文献
163.
MADELEINE C. THOMSON STEPHEN J. CONNOR MARTHA L. QUIÑONES MUSA JAWARA JAMES TODD BRIAN M. GREENWOOD 《Medical and veterinary entomology》1995,9(4):413-419
ovement of mosquitoes belonging to the Anopheles gambiae complex (mixed wild populations of An.arabiensis, An.gambiae and An.melas ) between three neighbouring rural villages in The Gambia was investigated by mark-release-recapture. A total of 12,872 mosquitoes were collected in bednets, marked with a magenta fluorescent powder and released over a 15-day period in one of the villages. A further 15,507 mosquitoes were collected in exit traps, marked with a yellow powder and released over the same period. Mosquitoes were captured daily in all three villages using pyrethrum spray catches, as well as bednet and exit trap catches. The catching period extended for 6 days after the last day of release.
Of the mosquitoes released, 372 (1.3%) were recaptured 2–21 days later. Of these recaptures, 272 were caught in the release village, and 98 were caught in other villages situated 1–1.4 km away. The 'movement index' between villages was calculated as 17.2% (12.2–22.4% confidence limits) for mosquitoes released after feeding and 20.1% (14.7–25.3%) for those released unfed.
These results suggest that movement of mosquitoes between neighbouring villages in The Gambia seriously affects the entomological evaluation of pyrethroid-impregnated bednet programmes in areas where treated and untreated villages are interspersed. 相似文献
Of the mosquitoes released, 372 (1.3%) were recaptured 2–21 days later. Of these recaptures, 272 were caught in the release village, and 98 were caught in other villages situated 1–1.4 km away. The 'movement index' between villages was calculated as 17.2% (12.2–22.4% confidence limits) for mosquitoes released after feeding and 20.1% (14.7–25.3%) for those released unfed.
These results suggest that movement of mosquitoes between neighbouring villages in The Gambia seriously affects the entomological evaluation of pyrethroid-impregnated bednet programmes in areas where treated and untreated villages are interspersed. 相似文献
164.
I. Yu. Malyshev E. B. Manukhina V. D. Mikoyan L. N. Kubrina A. F. Vanin 《FEBS letters》1995,370(3):159
Heat shock potentiated the nitric oxide production (EPR assay) in the liver, kidney, heart, spleen, intestine, and brain. The heat shock-induced sharp transient increase in the rate of nitric oxide production preceded the accumulation of heat shock proteins (HSP70) (Western blot analysis) as measured in the heart and liver. In all organs the nitric oxide formation was completely blocked by the NO-synthase inhibitor
(L-NNA). L-NNA also markedly attenuated the heat shock-induced accumulation of HSP70. The results suggests that nitric oxide is involved in the heat shock-induced activation of HSP70 synthesis. 相似文献
165.
In order to investigate the effect of the Pt(II) ion on the stacking interaction between tryptophan and a guanine base, the quenching of Trp fluorescence was monitored for some systems in the absence and presence of the metal ion, and the association constants were obtained by the analysis of Eadie-Hofstee plots. All spectral data suggested that the stacking interaction is enhanced by the Pt(II) coordination to the guanine N7 atom. The result indicates the importance of the metal ion as a bookmark in the specific recognition of a nucleic acid base by an aromatic amino acid residue. 相似文献
166.
During the production by mammalian cells of recombinant factor VIII from which the B domain was deleted (rFVIII), proteolytic
cleavages in the C-terminal part of the heavy chain were observed (Kjalke et al., 1995). By radioactive pulse labelling it
was investigated whether the cleavages took place inside the cells during protein synthesis or after release in the medium.
The rFVIII-producing CHO (Chinese hamster ovary) cells were cultured in the presence of 35S-methionine and then the cell lysate and the conditioned media were immunoprecipitated and analyzed by electrophoresis. By
pulse labelling and chasing for various time periods, it was shown that the cleavages only took place after secretion of the
protein from the cells. Adding cell lysate to uncleaved rFVIII caused cleavage of the heavy chain, as seen by loss of binding
to a monoclonal antibody specific for intact rFVIII, indicating that the cleavage was performed by proteinase(s) released
from the lysed cells. By incubating intact rFVIII with the multicatalytic proteinase (proteasome) present in cytoplasm and
nucleus of eukaryotic cells, loss of binding to the monoclonal antibody was observed. This indicates that the multicatalytic
proteinase, released from lysed rFVIII producing cells, could be responsible for the cleavage of rFVIII. Among several protease
inhibitors tested, only bacitracin was found to diminish the extent of cleavage. Phosphatidylserine also protected rFVIII
against cleavage, probably by binding to rFVIII.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
167.
The power of heteronuclear NMR spectroscopy to study macromoleculesand their complexes has been amply demonstrated over the last decade. Theobstacle to routinely applying these techniques to the study of DNA has beenthe synthesis of 13C,15N-labeled DNA. Here wepresent a simple and efficient method to generate isotope-labeled DNA forNMR studies that is as easy as that for isotope labeling of RNA. The methodwas used to synthesize a uniformly13 C,15N-labeled 32-nucleotide DNA that binds tohuman basic fibroblast growth factor with high affinity and specificity.Isotope-edited experiments were applied to the13 C,15N-labeled DNA bound to unlabeled protein,and the 13 C,15N-labeled DNA was also examined incomplex with 15N-labeled protein. The NMR experiments showthat the DNA adopts a well-defined stable structure when bound to theprotein, and illustrate the potential of13 C,15N-labeled DNA for structural studies ofDNA–protein complexes. 相似文献
168.
S. Aime Mauro Botta S. Geninatti Crich Giovanni B. Giovenzana Roberto Pagliarin Maurizio Piccinini Massimo Sisti Enzo Terreno 《Journal of biological inorganic chemistry》1997,2(4):470-479
A novel heptacoordinating ligand consisting of a thirteen-membered tetraazamacrocycle containing the pyridine ring and bearing
three methylenephosphonate groups (PCTP-[13]) has been synthesized. Its Gd(III) complex displays a remarkably high longitudinal
water proton relaxivity (7.7 mM–1 s–1 at 25 °C, 20 MHz and pH 7.5) which has been accounted for in terms of contributions arising from (1) one water molecule
bound to the metal ion, (2) hydrogen-bonded water molecules in the second coordination sphere, or (3) water molecules diffusing
near the paramagnetic chelate. Variable-temperature 17O-NMR transverse relaxation data indicate that the residence lifetime of the metal-bound water molecule is very short (8.0 ns
at 25 °C) with respect to the Gd(III) complexes currently considered as contrast agents for magnetic resonance imaging. Furthermore,
GdPCTP-[13] interacts with human serum albumin (HSA), likely through electrostatic forces. By comparing water proton relaxivity
data for the GdPCTP-[13]-HSA adduct, measured as a function of temperature and magnetic field strength, with those for the
analogous adduct with GdDOTP (a twelve-membered tetraaza macrocyclic tetramethylenephosphonate complex lacking a metal-bound
water molecule), it has been possible to propose a general picture accounting for the main determinants of the relaxation
enhancement observed when a paramagnetic Gd(III) complex is bound to HSA. Basically, the relaxation enhancement in these systems
arises from (1) water molecules in the hydration shell of the macromolecule and protein exchangeable protons which lie close
to the interaction site of the paramagnetic complex and (2) the metal bound water molecule(s). As far as the latter contribution
is concerned, the interaction with the protein causes an elongation of the residence lifetime of the metal-bound water molecule,
which limits, to some extent, the potential relaxivity enhancement expected upon the binding of the paramagnetic complex to
HSA.
Received: 27 January 1997 / Accepted: 12 May 1997 相似文献
169.
Susanne Metzger Andrea Erxleben B. Lippert 《Journal of biological inorganic chemistry》1997,2(2):256-264
The analogy between H-bonded nucleobase pairs and their metalated analogues is extended to the hemiprotonated pair of 7,9-dimethylguanine
(7,9-DimeG) and the Watson-Crick and reversed Watson-Crick pair between 7,9-dimethylguaninium (7,9-DimeGH+) and 1-methylcytosine (1-MeC). The crystal structure analyses of two model compounds, trans–[Pt(CH3NH2)2(7,9-DimeG-N1)2](NO3)2 (1) and trans–[Pt(NH3)2(1-MeC-N3)(7, 9-DimeG-N1)](PF6)2· 2.5 H2O (3a) are reported. Pt binding is through N1 of 7,9-DimeG and N3 of 1-MeC. In solution, 3a exists in a mixture with Watson-Crick and reversed Watson-Crick arrangements of the two bases, depending on solvent, concentration
and anions.
Received: 16 October 1996 / Accepted: 27 January 1997 相似文献
170.
Summary The oxidation of cysteine (RSH) has been studied by using O2, ferricytochrome c (Cyt c) and nitro blue tetrazolium (NBT) as electron acceptors. The addition of 200M CuII to a solution of 2mM cysteine, pH 7.4, produces an absorbance with a peak at 260 nm and a shoulder at 300 nm. Generation of a cuprous bis-cysteine complex (RS-CuI-SR) is responsible for this absorbance. In the absence of O2 the absorbance is stable for long time while in the presence of air it vanishes slowly only when the cysteine excess is consumed. The neocuproine assay and the EPR analysis show that the metal remains reduced in the course of the oxidation of cysteine returning to the oxidised form at the end of reaction when all RSH has been oxidised to RSSR. Addition of CuII enhances the reduction rate of Cyt c and of NBT by cysteine also under anaerobiosis indicating the occurrence of a direct reduction of the acceptor by the complex. It is concluded that the cuprous bis-cysteine complex (RS-CuI-SR) is the catalytic species involved in the oxidation of cysteine. The novel finding of the stability of the complex together with the metal remaining in the reduced form during the oxidation suggest sulfur as the electron donor in the place of the metal ion.Abbreviations RSH
cysteine
- RS–
cysteine in the thiolate form
- RS·
thiyl radical of cysteine
- RSSR
cystine
- Cyt c
cytochrome c
- SOD
superoxide dismutase
- NBT
nitro blue tetrazolium
- NBF
nitro blue formazan
- DTNB
5,5-dithiobis-2-nitrobenzoic acid
- DTPA
diethylenetriaminepentaacetic acid
Dedicated to prof. A. Ballio ob the occasion of his 75th birthday. 相似文献