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81.
Bennett T. Farmer II Luciano Müller Edward P. Nikonowicz Arthur Pardi 《Journal of biomolecular NMR》1994,4(1):129-133
Summary A set of three 3D (1H, 13C, 15N) triple-resonance correlation experiments has been designed to provide H1-H8 intraresidue sugar-to-base correlations in purines in an unambiguous and efficient manner. Together, the HsCsNb, HsCs(N)bCb, and HbNbCb experiments correlate the H1 sugar proton to the H8 proton of the attached base by means of the {H1, C1, N9, C8, H8} heteronuclear scalar coupling network. The assignment strategy presented here allows for unambiguous H1-H8 intraresidue correlations, provided that no two purines have both the same H1 and C1 chemical shifts and the same C8 and N9 chemical shifts. These experiments have yielded H1-H8 intraresidue sugar-to-base correlations for all five guanosines in the [13C, 15N] isotopically labeled RNA duplex r(GGCGCUUGCGUC)2. 相似文献
82.
83.
Summary A novel protocol for isotopically labeling bacterially expressed proteins is presented. This method circumvents problems related to poor cell growth, commonly associated with the use of minimal labeled media, and problems with protein induction encountered, less commonly, when using enriched labeled media. The method involves initially growing the bacterial cells to high optical density in a commercially available enriched labeled medium. Following a suitable growth period, the cells are transferred to a different (minimal) labeled medium, appropriate for induction. The method is demonstrated using the protein melanoma growth stimulating activity (MGSA). 相似文献
84.
Summary A 3D triple resonance experiment has been designed to provide intraresidual and sequential correlations between amide nitrogens and -carbons in uniformly 13C15N-labeled proteins. In-phase 13C magnetization is transferred to the aliphatic side-chain protons via the side-chain carbons using a CC-TOCSY mixing sequence. Thus, the experiment alleviates the resonance assignment process by providing information about the amino acid type as well as establishing sequential connectivities. Leaving the carbonyl spins untouched throughout the transfer from 13C to 1H leads to E.COSY-type cross peaks, from which the 3JH
co coupling constants can be evaluated. The pulse sequence is applied to oxidized Desulfovibrio vulgaris flavodoxin. 相似文献
85.
Nuclear magnetic resonance studies of the location and function of plant nutrients in vivo 总被引:2,自引:1,他引:1
The cytoplasmic and vacuolar pools of ammonium, inorganic phosphate and potassium can be studied non-invasively in plant tissues using high resolution nuclear magnetic resonance spectroscopy. The techniques that allow these pools to be discriminated in vivo are described and their application to plants is reviewed with reference to the phosphorus, nitrogen and potassium nutrition of root tissues. 相似文献
86.
Josefa Salgado Jose Villalain Juan C. Gómez-Fernández 《European biophysics journal : EBJ》1993,22(2):151-155
-Tocopherol, ubiquinone-10 and ubiquinol-10 have been studied by high resolution magic angle samples spinning 13C-nuclear magnetic resonance in egg yolk phosphatidylcholine multilamellar vesicles model membranes in order to assess their location and the induced perturbations on this model system. -Tocopherol is placed in such a position that it is in close contact with the head group of the phospholipid and exposed to the solvent. In this position it significantly perturbs the phospholipid head group, the 5a-CH3 and the 7a-CH3 groups being the closest parts to the membrane surface. On the other hand, ubiquinol-10 perturbs the membrane surface more than ubiquinone-10, but neither compound significantly changed the phospholipid head group conformation. 相似文献
87.
NMR imaging of root water distribution in intact Vicia faba L plants in elevated atmospheric CO2 总被引:1,自引:1,他引:0
The effect of elevated atmospheric CO2 on water distribution in the intact roots of Vicia faba L. bean seedlings grown in natural soil was studied noninvasively with proton (1H) nuclear magnetic resonance (NMR) imaging. Exposure of 24-d-old plants to atmospheric CO2-enriched air at 650 cm3 m?3 produced significant increases in water imaged in upper roots, hypogeal cotyledons and lower stems in response to a short-term drying-stress cycle. Above ground, drying produced negligible stem shrinkage and stomatal resistance was unchanged. In contrast, the same drying cycle caused significant depletion of water imaged in the same upper root structures in control plants subject to ambient CO2 (350 m3 m?3), and stem shrinkage and increased stomatal resistance. The results suggest that inhibition of transpiration caused by elevated CO2 does not necessarily result in attenuation of water transport from lower root structures. Inhibition of water loss from upper roots and lower stem in elevated CO2 environments may be a mitigating factor in assessing deleterious effects of greenhouse changes on crops during periods of dry climate. 相似文献
88.
Ronnitte S. Badar-Goffer Nicola M. Thatcher Peter G. Morris Herman S. Bachelard 《Journal of neurochemistry》1993,61(6):2207-2214
(1) The energy state and free intracellular calcium concentration ([Ca2±i) of super-fused cortical slices were measured in moderate hypoxia (~65 μM O2), in mild hypoglycaemia (0.5 mM glucose), and in combinations of the two insults using 19F and 31P NMR spectroscopy. (2) Neither hypoxia nor hypoglycaemia alone caused any significant change in [Ca2±i. Hypoxia caused a 40% fall in phosphocreatine (PCr) content but not in ATP level, and hypoglycaemia produced a slight fall in both (as expected from previous studies). These changes in the energy state recovered on return to control conditions. (3) A combined sequential insult (hypoxia, followed by hypoxia plus hypoglycaemia) produced a 100% increase in [Ca2±, and a decrease in PCr level to ~25% of control. The reverse combined sequential insult (hypoglycaemia, followed by hypoglycaemia plus hypoxia) had the same effect. On return to control conditions there was some decrease in [Ca2±i and a small increase in PCr content, but neither recovered to control levels. (4) Exposure of the tissue to the combined simultaneous insult (hypoxia plus hypoglycaemia) immediately after the control spectra had been recorded resulted in a fivefold increase in [Ca2±i and a similar decrease in PCr level to 20–25% of control. There was little if any change of [Ca2±i or PCr level on return to control conditions. (5) These results are discussed in terms of metabolic adaptation of some but not all of the cortical cells to the single type of insult, which renders the tissues less vulnerable to the combined insult. 相似文献
89.
Abstract: Ischemia-induced changes in 31 P NMR relaxation were examined in 16 piglets. NMR spectra were acquired under control conditions and during complete cerebral ischemia induced via cardiac arrest. Changes in T 1 were assessed directly in six animals during control conditions and after 30–45 min of complete ischemia when changes in brain P1 levels had reached a plateau. The T 1 for P1 did not change, i.e., 2.3 ± 0.5 s during control conditions versus 2.4 ± 1.0 s during ischemia. To evaluate phosphocreatine and ATP, two types of spectra, with a long (25-s) or short (1-s) interpulse delay time, were collected during the first 10 min of ischemia (n = 10). Both types of spectra showed the same time course of changes in phosphocreatine and ATP levels, implying that the T 1 relaxation times do not change during ischemia. There were no changes in the linewidths of phosphocreatine, ATP, or P1 during ischemia, implying that the T *2 values remain constant. Our results suggest that the 31 P T 1 and T *2 for phosphocreatine, Pi , and ATP do not change during ischemia, and therefore changes in 31 P NMR peak intensity accurately reflect changes in metabolite concentrations. 相似文献
90.
Comparison of glucose fermentation by suspended and gel-entrapped yeast cells: An in vivo nuclear magnetic resonance study 总被引:1,自引:0,他引:1
Phosphorus-31 nuclear magnetic resonance ((31)P NMR) was used to compare the anaerobic metabolism of glucose by suspended and gel-entrapped Saccharomyces bayanus cells. The fermentation of glucose was carried out in a reaction system with continuous circulation through the NMR sample tube. The intracellular pH and the levels of some phosphorylated compounds were the levels of some phosphorylated compounds were noninvasively monitored by (31)P NMR while glucose, fermentation products, and biomass were determined by analytic techniques comparisons showed that no significant differences are observed in the relative concentrations in the spectra, but distinct profiles for the variation of both intracellular and extracellular pH are found. The internal pH of immobilized cells is maintained at a constant value throughout the fermentation as opposed to freely suspended cells for which a steady decrease in the internal pH occurs. A faster and stronger acidification is also observed in the external medium of the assays with suspended cells. Furthermore, higher yields for ethanol and biomass production and lower yields of fermentation by-products are obtained with immobilized cells. It is concluded that the higher intracellular pH achieved in the presence of the gel matrix had a regulatory effect on the metabolism which favored the ethanol production pathway. (c) 1993 John Wiley & Sons, Inc. 相似文献