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11.
Analysis of the energy metabolism after incubation of Saccharomyces cerevisiae with sulfite or nitrite 总被引:1,自引:0,他引:1
After addition of 5 mM sulfite or nitrite to glucose-metabolizing cells of Saccharomyces cerevisiae a rapid decrease of the ATP content and an inversely proportional increase in the level of inorganic phosphate was observed. The concentration of ADP shows only small and transient changes. Cells of the yeast mutant pet 936, lacking mitochondrial F1ATPase, after addition of 5 mM sulfite or nitrite exhibit changes in ATP, ADP and inorganic phosphate very similar to those observed in wild type cells. They key enzyme of glucose degradation, glyceraldehyde-3-phosphate dehydrogenase was previously shown to be the most sulfiteor nitrite-sensitive enzyme of the glycolytic pathway. This enzyme shows the same sensitivity to sulfite or nitrite in cells of the mutant pet 936 as in wild type cells. It is concluded that the effects of sulfite or nitrite on ATP, ADP and inorganic phosphate are the result of inhibition of glyceraldehyde-3-phosphate dehydrogenase and not of inhibition of phosphorylation processes in the mitochondria. Levels of GTP, UTP and CTP show parallel changes to ATP. This is explained by the presence of very active nucleoside monophosphate kinases which cause a rapid exchange between the nucleoside phosphates. The effects of the sudden inhibition of glucose degradation by sulfite or nitrite on levels of ATP, ADP and inorganic phosphate are discussed in terms of the theory of Lynen (1942) on compensating phosphorylation and dephosphorylation in steady state glucose metabolizing yeast.Abbreviations ATP
adenosine triphosphate
- ADP
adenosine diphosphate
- AMP
adenosine monophosphate
- Pi
inorganic orthophosphate
Dedicated to Prof. Dr. Hans Grisebach on the occasion of his sixtieth birthday 相似文献
12.
Evidence is presented here that axenic cultures of Nostoc spp., Aphanocapsa (PCC 6308), and Aphanocapsa (PCC 6714) but not Anacystis nidulans R-2 (PCC 7942) produce N2O and ammonia when grown on nitrite. The data suggest that the cyanobacteria produce N2O by nitrite reduction to ammonia.Nonstandard abbreviations DCMU
3-(3,4-dichlorophenyl)-1,1-dimethyl urea
- NIR
nitrite reductase 相似文献
13.
Plastids were separated from extracts of pea (Pisum sativum L.) roots by sucrose-density-gradient centrifugation. The incubation of roots of intact pea seedlings in solutions containing 10 mM KNO3 resulted in increased plastid activity of nitrite reductase and to a lesser extent glutamine synthetase. There were also substantial increases in the activity of glucose-6-phosphate and 6-phosphogluconate dehydrogenases. No other plastid-located enzymes of nitrate assimilation or carbohydrate oxidation showed evidence of increased activity in response to the induction of nitrate assimilation. Studies with [1-14C]-and [6-14C]glucose indicated that there was an increased flow of carbon through the plastid-located pentose-phosphate pathway concurrent with the induction of nitrate assimilation. It is suggested that there is a close interaction through the supply and demand for reductant between the pathway of nitrite assimilation and the pentose-phosphate pathway located in the plastid. 相似文献
14.
Nitrogen-starved sunflower plants (Helianthus annuus L. cv. Peredovic) cannot absorb NO
3
–
or NO
2
–
upon initial exposure to these anions. Ability of the plants to take up NO
3
–
and NO
2
–
at high rates from the beginning was induced by a pretreatment with NO
3
–
. Nitrite also acted as inducer of the NO
2
–
-uptake system. The presence of cycloheximide during NO
3
–
-pretreatment prevented the subsequent uptake of NO
3
–
and NO
2
–
, indicating that both uptake systems are synthesized de novo when plants are exposed to NO
3
–
. Cycloheximide also suppressed nitrate-reductase (EC 1.6.6.1) and nitrite-reductase (EC 1.7.7.1) activities in the roots. The sulfhydryl-group reagent N-ethylmaleimide greatly inhibited the uptake of NO
3
–
and NO
2
–
. Likewise, N-ethylmaleimide promoted in vivo the inactivation of nitrate reductase without affecting nitrite-reductase activity. Rates of NO
3
–
and NO
2
–
uptake as a function of external anion concentration exhibited saturation kinetics. The calculated Km values for NO
3
–
and NO
2
–
uptake were 45 and 23 M, respectively. Rates of NO
3
–
uptake were four to six times higher than NO
3
–
-reduction rates in roots. In contrast, NO
2
–
-uptake rates, found to be very similar to NO
3
–
-uptake rates, were much lower (about 30 times) than NO
2
–
-reduction rates. Removal of oxygen from the external solution drastically suppressed NO
3
–
and NO
2
–
uptake without affecting their reduction. Uptake and reduction were also differentially affected by pH. The results demonstrate that uptake of NO
3
–
and NO
2
–
into sunflower plants is mediated by energy-dependent inducible-transport systems distinguishable from the respective enzymatic reducing systems.Abbreviations CHI
cycloheximide
- NEM
N-ethylmaleimide
- NiR
nitrite reductase
- NR
nitrate reductase
-
pHME
p-hydroxymercuribenzoate
This research was supported by grant PB86-0232 from the Dirección General de Investigatión Científica y Técnica (Spain). One of us (E.A.) thanks the Consejeria de Educación y Ciencia de la Junta de Andalucia for the tenure of a fellowship. We thank Miss G. Alcalá and Miss C. Santos for their valuable technical and secretarial assistance. 相似文献
15.
It has been suggested that two groups ofEscherichia coli genes, theccm genes located in the 47-min region and thenrfEFG genes in the 92-min region of the chromosome, are involved in cytochromec biosynthesis during anaerobic growth. The involvement of the products of these genes in cytochromec synthesis, assembly and secretion has now been investigated. Despite their similarity to other bacterial cytochromec assembly proteins, NrfE, F and G were found not to be required for the biosynthesis of any of thec-type cytochromes inE. coli. Furthermore, these proteins were not required for the secretion of the periplasmic cytochromes, cytochromec
550 and cytochromec
552, or for the correct targeting of the NapC and NrfB cytochromes to the cytoplasmic membrane. NrfE and NrfG are required for formate-dependent nitrite reduction (the Nrf pathway), which involves at least twoc-type cytochromes, cytochromec
552 and NrfB, but NrfF is not essential for this pathway. Genes similar tonrfE, nrfF andnrfG are present in theE. coli nap-ccm locus at minute 47. CcmF is similar to NrfE, the N-terminal region of CcmH is similar to NrfF and the C-terminal portion of CcmH is similar to NrfG. In contrast to NrfF, the N-terminal, NrfF-like portion of CcmH is essential for the synthesis of allc-type cytochromes. Conversely, the NrfG-like C-terminal region of CcmH is not essential for cytochromec biosynthesis. The data are consistent with proposals from this and other laboratories that CcmF and CcmH form part of a haem lyase complex required to attach haemc to C-X-X-C-H haem-binding domains. In contrast, NrfE and NrfG are proposed to fulfill a more specialised role in the assembly of the formate-dependent nitrite reductase. 相似文献
16.
Sheldon Milstien Naoki Sakai Bruce J. Brew †Charles Krieger ‡James H. Vickers §Kuniaki Saito §Melvyn P. Heyes 《Journal of neurochemistry》1994,63(3):1178-1180
Abstract: Nitric oxide has been proposed to mediate cytotoxic effects in inflammatory diseases. To investigate the possibility that overproduction of nitric oxide might play a role in the neuropathology of inflammatory and noninflammatory neurological diseases, we compared levels of the markers of nitric oxide, nitrite plus nitrate, in the CSF of controls with those in patients with various neurologic diseases, including Huntington's and Alzheimer's disease, amyotrophic lateral sclerosis, and HIV infection. We found that there were no significant increases in the CSF levels of these nitric oxide metabolites, even in patients infected with HIV or in monkeys infected with poliovirus, both of which have significantly elevated levels of the neurotoxin quinolinic acid and the marker of macrophage activation, neopterin. However, CSF quinolinic acid, neopterin, and nitrite/nitrate levels were significantly increased in a small group of patients with bacterial and viral meningitis. 相似文献
17.
T. M. Hennessey L. E. Frego J. T. Francis 《Journal of comparative physiology. A, Neuroethology, sensory, neural, and behavioral physiology》1994,175(5):655-665
Paramecium is a valuable eukaryotic model system for studying chemosensory transduction, adaptation and cellular sensory integration. While millimolar amounts of many attractants hyperpolarize and cause faster forward swimming, oxidants are repellents that depolarize and cause backward swimming at micromolar concentrations. The non-permeant oxidants cytochrome c, nitro blue tetrazolium and ferricyanide are repellents with half maximal concentrations of 0.4 M, 2.2 M and 100 M respectively. In vivo reductase activities follow the same order of potencies. The concentration dependence of the cytochrome c reductase activity is well correlated with cytochrome c-induced depolarizations. This suggests that plasma membrane reduction of external cytochrome c is electrogenic, causing membrane depolarization and chemorepulsion. The reductase activity also appears to be voltage dependent. Depolarization by either K+, Na+, Ca+ or Mg+ correlates with inhibition of both in vivo reductase activities and cytochrome c-induced membrane potential changes. These responses were also seen in deciliated cells, showing that the body plasma membrane is sufficient for the response. Both chloroquine and diphenyleneiodonium inhibited reductase activities but only at unusually high concentrations. This activity showed no pH dependence in the physiological range. We propose that a plasma membrane bound NADPH-dependent reductase controls oxidant-induced depolarizations and consequent chemorepulsion.Abbreviations
bmv
Body plasma membrane vesicles
-
BPS
Bathophenanthroline disulfonate
-
cAMP
Cyclic adenosine monophosphate
-
cmv
Ciliary membrane vesicles
-
cyt c
Cytochrome c
-
DPI
Diphenyleneiodonium
-
EC
50
Concentration for 50% effectiveness
-
FeCN
Ferricyanide [Fe(CN)6–3]
-
FeEDTA
Ethylenediaminetetracetic acid (ferric-sodium salt)
-
GTP
Guanosine 5-triphosphate
-
KCN
Potassium cyanide
-
mM
Millimolar
-
MOPS
3-(N-morpholino) propanesulfonic acid
-
mV
Millivolts
-
NADH
Nicotinamide adenine dinucleotide (reduced form)
-
NADPH
Nicotinamide adenine dinucleotide phosphate (reduced form)
-
NBT
Nitro blue tetrazolium
-
nm
Nanometer
-
pCMB
p-Chloromercuribenzoate
-
PMA
Phorbol 12-myristate 13-acetate
-
s.d.
Standard deviation
-
SOD
Superoxide dismutase
-
Tris
Tris(hydroxymethyl)aminomethane
-
M
Micromolar 相似文献
18.
Jocelyne Kronenberger Andrée Lepingle Michel Caboche Hervé Vaucheret 《Molecular & general genetics : MGG》1993,236(2-3):203-208
Summary Three tobacco nitrite reductase (NiR) cDNA clones were isolated using spinach NiR cDNA as a probe. Sequence analysis and Southern blot hybridization revealed four genes in tobacco. Two of these genes presumably derived from the ancestral species Nicotiana tomentosiformis, the other two from the ancestor N. sylvestris. Northern blot analysis showed that one gene from each ancestral genome was expressed predominantly in leaves, whilst RNA from the other was detected mostly in roots. The accumulation of both leaf and root NiR mRNAs was induced by nitrate and repressed by nitrate- or ammonium-derived metabolites. In addition, the expression of the root NiR gene was detectable in leaves of a tobacco nitrate reductase (NR)-deficient mutant. Thus, the regulation of expression of tobacco NiR genes is comparable to the regulation of expression of barley NR genes. 相似文献
19.
The effect of nitrogen starvation on the NO3-dependent induction of nitrate reductase (NR) and nitrite reductases (NIR) has been investigated in the halophilic alga Dunaliella salina. When D. salina cells previously grown in a medium with NH 4 + as the only nitrogen source (NH 4 + -cells) were transferred into NO 3 ? medium, NR was induced in the light. In contrast, when cells previously grown in N-free medium were transferred into a medium containing NO 3 ? , NR was induced in light or in darkness. Nitrate-dependent NR induction, in darkness, in D. salina cells previously grown at a photon flux density of 500 umol · m?2 s?1 was observed after 4 h preculture in N-free medium, whilst in cells grown at 100 umol · m?2 s?1 NR induction was observed after 7–8 h. An inhibitor of mRNA synthesis (6-methylpurine) did not inhibit NO 3 ? -induced NR synthesis when the cells, previously grown in NH 4 + medium, were transferred into NO 3 ? medium (at time 0 h) after 4-h-N starvation. However, when 6-methylpurine was added simultaneously with the transfer of the cells from NH 4 + to NO 3 ? medium (at time 0 h), NO 3 ? induced NR synthesis was completely inhibited. The activity of NIR decreased in N-starved cells and the addition of NO 3 ? to those cells greatly stimulated NIR activity in the light. The ability to induce NR in darkness was observed when glutamine synthetase activity reached its maximal level during N starvation. Although cells grown in NO 3 ? medium exhibited high NR activity, only 0.33% of the total NR was found in intact chloroplasts. We suggest that the ability, to induce NR in darkness is dependent on the level of N starvation, and that NR in D. salina is located in the cytosol. Light seems to play an indirect regulatory role on NO 3 ? uptake and NR induction due to the expression of NR and NO 3 ? -transporter mRNAs. 相似文献
20.
Elisabetta Zennaro Ilaria Ciabatti Francesca Cutruzzola Rosanna D'Alessandro Maria Chiara Silvestrini 《FEMS microbiology letters》1993,109(2-3):243-250
Abstract The expression of nitrite reductase has been tested in a wild-type strain of Pseudomonas aeruginosa (Pao1) as a function of nitrate concentration under anaerobic and aerobic conditions. Very low levels of basal expression are shown under non-denitrifying conditions (i.e. absence of nitrate, in both aerobic and anaerobic conditions); anaerobiosis is not required for high levels of enzyme production in the presence of nitrate. A Pseudomonas aeruginosa strain, mutated in the nitrite reductase gene, has been obtained by gene replacement. This mutant, the first of this species described up to now, is unable to grow under anaerobic conditions in the presence of nitrate. The anaerobic growth can be restored by complementation with the wild-type gene. 相似文献