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1.
Nature ; 633(8029): 306-317, 2024 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-39261612

RESUMO

Radiocarbon (14C) is essential for creating chronologies to study the timings and drivers of pivotal events in human history and the Earth system over the past 55,000 years. It is also a fundamental proxy for investigating solar processes, including the potential of the Sun for extreme activity. Until now, fluctuations in past atmospheric 14C levels have limited the dating precision possible using radiocarbon. However, the discovery of solar super-storms known as extreme solar particle events (ESPEs) has driven a series of advances with the potential to transform the calendar-age precision of radiocarbon dating. Organic materials containing unique 14C ESPE signatures can now be dated to annual precision. In parallel, the search for further storms using high-precision annual 14C measurements has revealed fine-scaled variations that can be used to improve calendar-age precision, even in periods that lack ESPEs. Furthermore, the newly identified 14C fluctuations provide unprecedented insight into solar variability and the carbon cycle. Here, we review the current state of knowledge and share our insights into these rapidly developing, diverse research fields. We identify links between radiocarbon, archaeology, solar physics and Earth science to stimulate transdisciplinary collaboration, and we propose how researchers can take advantage of these recent developments.

2.
Ann Bot ; 118(3): 401-14, 2016 09.
Artigo em Inglês | MEDLINE | ID: mdl-27474507

RESUMO

BACKGROUNDS AND AIMS: Crops with reduced requirement for nitrogen (N) fertilizer would have substantial benefits in developed nations, while improving food security in developing nations. This study employs the functional structural plant model SimRoot to test the hypothesis that variation in the growth angles of axial roots of maize (Zea mays L.) is an important determinant of N capture. METHODS: Six phenotypes contrasting in axial root growth angles were modelled for 42 d at seven soil nitrate levels from 10 to 250 kg ha(-1) in a sand and a silt loam, and five precipitation regimes ranging from 0·5× to 1·5× of an ambient rainfall pattern. Model results were compared with soil N measurements of field sites with silt loam and loamy sand textures. KEY RESULTS: For optimal nitrate uptake, root foraging must coincide with nitrate availability in the soil profile, which depends on soil type and precipitation regime. The benefit of specific root architectures for efficient N uptake increases with decreasing soil N content, while the effect of soil type increases with increasing soil N level. Extreme root architectures are beneficial under extreme environmental conditions. Extremely shallow root systems perform well under reduced precipitation, but perform poorly with ambient and greater precipitation. Dimorphic phenotypes with normal or shallow seminal and very steep nodal roots performed well in all scenarios, and consistently outperformed the steep phenotypes. Nitrate uptake increased under reduced leaching conditions in the silt loam and with low precipitation. CONCLUSIONS: Results support the hypothesis that root growth angles are primary determinants of N acquisition in maize. With decreasing soil N status, optimal angles resulted in 15-50 % greater N acquisition over 42 d. Optimal root phenotypes for N capture varied with soil and precipitation regimes, suggesting that genetic selection for root phenotypes could be tailored to specific environments.


Assuntos
Nitrogênio/metabolismo , Raízes de Plantas/crescimento & desenvolvimento , Zea mays/crescimento & desenvolvimento , Produtos Agrícolas , Meio Ambiente , Fertilizantes , Modelos Teóricos , Nitratos/metabolismo , Raízes de Plantas/fisiologia , Solo/química , Zea mays/anatomia & histologia , Zea mays/fisiologia
3.
J Radioanal Nucl Chem ; 311(1): 409-418, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28111485

RESUMO

Molybdenum-99 is one of the most important radionuclides for medical diagnostics. In 2015, the International Atomic Energy Agency organized a round-robin exercise where the participants measured and calculated specific saturation activities achievable for the 98Mo(n,γ)99Mo reaction. This reaction is of interest as a means to locally, and on a small scale, produce 99Mo from natural molybdenum. The current paper summarises a set of experimental results and reviews the methodology for calculating the corresponding saturation activities. Activation by epithermal neutrons and also epithermal neutron self-shielding are found to be of high importance in this case.

4.
J Mol Biol ; 238(5): 857-9, 1994 May 20.
Artigo em Inglês | MEDLINE | ID: mdl-8182756

RESUMO

Single crystals of the lipase from Bacillus subtilis have been obtained using a mixture of polyethylene glycol 4000 and sodium sulphate solution as the precipitant. The crystals grow at room temperature in two to three weeks in the presence of n-octyl-beta-D-glucoside. They belong to the monoclinic space group C2 with a = 121.20 A, b = 93.19 A, c = 80.96 A, and beta = 110.67 degrees, with four protein molecules per asymmetric unit. The crystals diffract to at least 2.5 A resolution and are suitable for an X-ray structure analysis.


Assuntos
Bacillus subtilis/enzimologia , Lipase/química , Cristalização , Cristalografia por Raios X , Estrutura Molecular
5.
Appl Radiat Isot ; 62(3): 429-33, 2005 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-15607919

RESUMO

Neutron resonance capture analysis was applied to a bronze commemorative plaque from the West-African country Benin. By comparison with recently published element compositions of Benin memorial heads, the alloy of the plaque could be dated to the period 1725-1897 AD. In the analysis procedure, the object was not damaged, cleaned or altered, and very little long-lived radioactivity was induced.

6.
Gene ; 252(1-2): 71-82, 2000 Jul 11.
Artigo em Inglês | MEDLINE | ID: mdl-10903439

RESUMO

We established a tetracycline-regulated gene expression system that tightly controls expression of genes in Dictyostelium discoideum. The control elements are contained in two plasmid vectors, one being an integrated plasmid encoding a chimeric tetracycline-controlled transcriptional activator protein (tTA(s)(*)). The second component is an extrachromosomal plasmid harboring the gene of interest preceded by an inducible promoter. This promoter contains a tetracycline-responsive element, which is the binding site for tTA(s)(*). Tetracycline prevents tTA(s)(*) from binding to the tetracycline-responsive element, rendering the promoter virtually silent. In the absence of tetracycline, tTA(s)(*) binds to its target sequence and strongly induces gene expression. The kinetics of activation and repression of the system were monitored using luciferase as a reporter. The results reveal efficient inhibition of gene expression by low concentrations of tetracycline and an induction of gene expression by several orders of magnitude within a few hours after removal of tetracycline. Green fluorescent protein (GFP) provided information about the effects of modulation of the tetracycline concentration on gene expression, at the single cell level, using fluorescence activated cell sorting (FACS). We also report that not all cells in a clonal population express the reporter gene.


Assuntos
Dictyostelium/genética , Tetraciclina/farmacologia , Transativadores/efeitos dos fármacos , Actinas/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , DNA Recombinante/genética , Dictyostelium/citologia , Relação Dose-Resposta a Droga , Citometria de Fluxo , Regulação da Expressão Gênica/efeitos dos fármacos , Vetores Genéticos , Proteínas de Fluorescência Verde , Cinética , Luciferases/efeitos dos fármacos , Luciferases/genética , Luciferases/metabolismo , Proteínas Luminescentes/efeitos dos fármacos , Proteínas Luminescentes/genética , Proteínas Luminescentes/metabolismo , Dados de Sequência Molecular , Plasmídeos , Regiões Promotoras Genéticas/genética , Proteínas Recombinantes de Fusão/efeitos dos fármacos , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/metabolismo , Elementos de Resposta/genética , Transativadores/genética
7.
FEBS Lett ; 373(1): 10-2, 1995 Oct 02.
Artigo em Inglês | MEDLINE | ID: mdl-7589423

RESUMO

The outer membrane phospholipase A (OMPLA) of Escherichia coli is one of the few integral outer membrane proteins displaying enzymatic activity. It is encoded as a mature protein of 269 amino acids preceded by a signal sequence of 20 amino acids. There is no sequence homology with water-soluble lipases and phospholipases. Crystals of the mature enzyme were obtained at 22 degrees C from 24-28% (v/v) 2-methyl-2,4-pentanediol in Bis-Tris buffer, pH 5.9-6.0, with 1 mM calcium chloride and 1.5% (w/v) beta-octylglucoside. They have the symmetry of the trigonal spacegroup P3(1)21 (or P3(2)21) with cell dimensions of a = b = 79.6 A and c = 102.8 A (alpha = beta = 90 degrees, gamma = 120 degrees). Native crystals diffract to a resolution of 2.6 A.


Assuntos
Escherichia coli/enzimologia , Fosfolipases A/química , Sequência de Aminoácidos , Membrana Celular/enzimologia , Clonagem Molecular , Cristalização , Cristalografia por Raios X/métodos , Indicadores e Reagentes , Dados de Sequência Molecular , Fosfolipases A/biossíntese , Fosfolipases A/isolamento & purificação , Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/química , Proteínas Recombinantes/isolamento & purificação
8.
Mol Biochem Parasitol ; 55(1-2): 115-26, 1992 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-1435864

RESUMO

Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) activity was detected in two cell compartments of Leishmania mexicana promastigotes. These activities could be attributed to two different isoenzymes, one residing in glycosomes, the other in the cytosol. We have cloned and sequenced the genes for both isoenzymes. The glycosomal enzyme is encoded by two tandemly linked genes of identical sequence and contains features frequently found in glycosomal enzymes: the presence of peptide insertions, a small carboxy-terminal extension with a potential glycosomal targeting signal (-SKM) and an excess of positively charged residues (net charge +7). Only one open reading frame was detected for the cytosolic enzyme. The amino acid sequences of the two proteins are only 55% identical. We discuss some evolutionary aspects of the observed organization of the GAPDH genes in the Trypanosomatidae and the role of the two isoenzymes in the metabolism of these organisms. The possibility to develop GAPDH-specific inhibitors that will be effective against the enzyme of various parasitic members of this family is explored.


Assuntos
DNA de Protozoário/química , Gliceraldeído-3-Fosfato Desidrogenases/genética , Isoenzimas/genética , Leishmania mexicana/enzimologia , Sequência de Aminoácidos , Animais , Sequência de Bases , Southern Blotting , Clonagem Molecular , Códon/genética , Citosol/enzimologia , Digitonina , Ativação Enzimática , Glucosefosfato Desidrogenase/metabolismo , Gliceraldeído-3-Fosfato Desidrogenases/química , Gliceraldeído-3-Fosfato Desidrogenases/metabolismo , Hexoquinase/metabolismo , Isoenzimas/química , Leishmania mexicana/classificação , Leishmania mexicana/genética , Dados de Sequência Molecular , Fases de Leitura Aberta , Organelas/enzimologia , Filogenia , Sequências Repetitivas de Ácido Nucleico , Mapeamento por Restrição , Análise de Sequência de DNA
9.
J Nutr Health Aging ; 18(6): 601-7, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-24950151

RESUMO

OBJECTIVES: To assess the prevalence of (risk of) undernutrition in Dutch elder Parkinson's disease patients as well as it's risk factors. DESIGN: Observational cross-sectional study. SETTING: An outpatient clinic at the department Neurology of Medical Centre Leeuwarden, a large teaching hospital. PARTICIPANTS: 102 outpatients with Parkinson's disease aged 65 years and older were recruited. MEASUREMENTS: Data regarding various aspects of undernutrition including socio-demographic aspect, disease characterisitics, nutritional status, appetite and overall-physical and psychological functioning were collected. RESULTS: Undernutrition was diagnosed in 2.0% and 20.5% of the patients were categorized as being at risk of undernutrition. Care dependency and appetite were the two risk factors with the highest predictive value for an unfavorable nutritional status. CONCLUSION: Of Dutch elderly patients with Parkinson's Disease 22.5% had an unfavourable nutritional status. Dependency and appetite were the two risk factors with the highest predictive value fort his outcome. Because undernutrition can be regarded as a geriatric syndrome a comprehensive nutritional assessment should be done followed by nutritional interventions next to interventions focused on the risk factors. Further studies are needed to evaluate these interventions.


Assuntos
Avaliação Geriátrica , Desnutrição/epidemiologia , Estado Nutricional , Doença de Parkinson/epidemiologia , Idoso , Idoso de 80 Anos ou mais , Apetite , Estudos Transversais , Dependência Psicológica , Feminino , Hospitais de Ensino , Humanos , Masculino , Desnutrição/diagnóstico , Países Baixos/epidemiologia , Avaliação Nutricional , Doença de Parkinson/fisiopatologia , Doença de Parkinson/psicologia , Prevalência , Fatores de Risco
11.
Appl Radiat Isot ; 69(2): 320-6, 2011 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-20727777

RESUMO

High-precision measurements were conducted on the time evolution of gamma-ray count rates during reactor-on and reactor-off periods to investigate the possible influence of antineutrinos on nuclear decay. This experiment was triggered by a recent analysis (Jenkins et al., 2009) of long-term measurements suggesting a possible link to variations in nuclear decay rate and solar neutrino flux. The antineutrino flux during reactor-off periods is mainly due to geoneutrinos and four orders of magnitude lower than during reactor-on periods. No effects have been observed for the two branches in the decay of (152)Eu and the decay of (137)Cs, (54)Mn and (22)Na. The upper limit determined of the ratio Δλ/λ for (22)Na is (-1±2)×10(-4), and (54)Mn is (-1±4)×10(-4). In comparison to the interpretation of Jenkins et al. our measurements do not show any such effect to at least two orders of magnitude less. Hence either the hypothesis of Jenkins et al. is not true or else one of two rather unlikely possibilities must also be true: either the effect of neutrinos on ß(-) decay differs considerably from the effect of antineutrinos on ß(+) decay, or the effect of antineutrinos on ß(+) decay must be identical to their effect on ß(-) and electron-capture decay.

12.
Appl Radiat Isot ; 69(9): 1181-4, 2011 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-21550260

RESUMO

Neutron activation of Sm-152 offers a method of radiolabeling for the in vivo study of oral dose formulations by gamma scintigraphy. Reproducibility measurements are needed to ensure the robustness of clinical studies. 204 enteric-coated guaifenesin core tablets (10mg of Sm(2)O(3)) were irradiated by thermal neutrons to achieve 1 MBq at 48 h. Administered activities were 0.86±0.03 MBq. Good reproducibility (CV=3.5%) was observed over 24 weeks ensuring that volunteer doses were within the dose reference level of 0.8 mSv.


Assuntos
Guaifenesina/análogos & derivados , Administração Oral , Guaifenesina/administração & dosagem , Humanos , Marcação por Isótopo , Análise de Ativação de Nêutrons , Radioisótopos , Reprodutibilidade dos Testes , Samário
13.
Oecologia ; 142(3): 428-39, 2005 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-15526119

RESUMO

Knowledge of the interactions between organisms within trophic groups is important for an understanding of the role of biodiversity in ecosystem functioning. We hypothesised that interactions between bacterivorous nematodes of different life history strategies would affect nematode population development, bacterial community composition and activity, resulting in increased N mineralization. A microcosm experiment was conducted using three nematode species (Bursilla monhystera, Acrobeloides nanus and Plectus parvus). All the nematode species interacted with each other, but the nature and effects of these interactions depended on the specific species combination. The interaction between B. monhystera and A. nanus was asymmetrically competitive (0,-), whereas that between B. monhystera and P. parvus, and also A. nanus and P. parvus was contramensal (+, -). The interaction that affected microcosm properties the most was the interaction between B. monhystera and P. parvus. This interaction affected the bacterial community composition, increased the bacterial biomass and increased soil N mineralization. B. monhystera and P. parvus have the most different life history strategies, whereas A. nanus has a life history strategy intermediate to those of B. monhystera and P. parvus. We suggest that the difference in life history strategies between species of the same trophic group is of importance for their communal effect on soil ecosystem processes. Our results support the idiosyncrasy hypothesis on the role of biodiversity in ecosystem functioning.


Assuntos
Bactérias/crescimento & desenvolvimento , Nematoides/fisiologia , Nitrogênio/análise , Microbiologia do Solo , Solo/análise , Animais , Bactérias/classificação , Ecossistema , Nematoides/classificação , Raízes de Plantas/microbiologia
14.
Biochemistry ; 26(18): 5796-803, 1987 Sep 08.
Artigo em Inglês | MEDLINE | ID: mdl-3314996

RESUMO

The mannitol-specific enzyme II (EII), purified free of phospholipid, exhibits a concentration dependence in its specific activity with P-HPr and mannitol as the donor and acceptor substrates, respectively. This concentration dependence, previously observed only in the case of mannitol----mannitol phosphate exchange reaction, indicates that an oligomeric form of the enzyme is responsible for catalyzing the phosphorylation reaction (P-HPr + mannitol----mannitol-P + HPr) as well as the exchange reaction. Kinetic analysis revealed that the monomeric enzyme has a much lower specific activity than the associated species. The specific activity can be increased by raising the steady-state level of phosphorylation of EII and also by adding phospholipid, demonstrating that phosphorylation and the binding of phospholipid facilitate the association process. Kinetic measurements and fluorescence energy transfer measurements demonstrate a strong preference of EII for phospholipids with specific head group and fatty acid composition.


Assuntos
Escherichia coli/enzimologia , Sistema Fosfotransferase de Açúcar do Fosfoenolpiruvato/metabolismo , Fosfolipídeos/metabolismo , Membrana Celular/enzimologia , Proteínas de Escherichia coli , Cinética , Proteínas de Transporte de Monossacarídeos , Fosfatidilgliceróis/farmacologia , Sistema Fosfotransferase de Açúcar do Fosfoenolpiruvato/isolamento & purificação , Fosforilação , Ligação Proteica
15.
Biol Cybern ; 58(3): 141-8, 1988.
Artigo em Inglês | MEDLINE | ID: mdl-3358949

RESUMO

Non-visually triggered arm movements over a horizontal table at shoulder height were analysed by an Information Theory approach according to a method suggested by Sakitt et al. (1983) and Sakitt (1980). The movement track was along the subject's median line and was indicated by a vertical metal ridge fixed to the table. The observer passively moved the subject's left index finger along the left side of the ridge to the target position. The blindfolded subject then had to move his right index finger along the right side of the ridge to match the left finger position. Direct contact between the two fingers was prevented by the ridge. We compared our results, which involve the transmission of information through the arm and shoulder joints of both arms, with those of Sakitt et al. which involved just one elbow joint. We supplemented our experimental results with simulations and show that the value for the transmitted information, obtained using the method of analysis suggested by Sakitt et al., is very dependent upon the number of trials, and number and spacing of the targets. Sakitt et al. suggest that the Information Theory approach permits easy comparison between different tasks and different observers. Our results suggest that comparisons should be made with caution.


Assuntos
Teoria da Informação , Movimento , Propriocepção , Adulto , Feminino , Dedos , Humanos , Masculino , Matemática , Pessoa de Meia-Idade
16.
Anal Chem ; 69(13): 2247-50, 1997 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-21639356

RESUMO

The feasibility of quantitative instrumental neutron activation analysis (INAA) of samples in the kilogram range without internal standardization has been demonstrated by Overwater et al. (Anal. Chem. 1996, 68, 341). In their studies, however, they demonstrated only the agreement between the "corrected" γ ray spectrum of homogeneous large samples and that of small samples of the same material. In this paper, the k(0) calibration of the IRI facilities for large samples is described, and, this time in terms of (trace) element concentrations, some of Overwater's results for homogeneous materials are presented again, as well as results obtained from inhomogeneous materials and subsamples thereof. It is concluded that large-sample INAA can be as accurate as ordinary INAA, even when applied to inhomogeneous materials.

17.
Biochemistry ; 23(21): 4934-9, 1984 Oct 09.
Artigo em Inglês | MEDLINE | ID: mdl-6437444

RESUMO

Purified mannitol-specific enzyme II (EIImtl), in the presence of the detergent Lubrol, catalyzes the phosphorylation of mannitol from P-HPr via a classical ping-pong mechanism involving the participation of a phosphorylated EIImtl intermediate. This intermediate has been demonstrated by using radioactive phosphoenolpyruvate. Upon addition of mannitol, at least 80% of the enzyme-bound phosphoryl groups can be converted to mannitol 1-phosphate. The EIImtl concentration dependence of the exchange reaction indicates that self-association is a prerequisite for catalytic activity. The self-association can be achieved by increasing the EIImtl concentration or at low concentrations of EIImtl by adding HPr or bovine serum albumin. The equilibrium is shifted toward the dissociated form by mannitol 1-phosphate, resulting in a mannitol 1-phosphate induced inhibition. Mannitol does not affect the association state of the enzyme. Both mannitol and mannitol 1-phosphate also act as classical substrate inhibitors. The apparent Ki of each compound, however, is approximately equal to its apparent Km, suggesting that mannitol and mannitol 1-phosphate bind at the same site on EIImtl. Due to strong inhibition provided by mannitol and mannitol 1-phosphate in the exchange reaction, the kinetics of this reaction cannot be used to determine whether the reaction proceeds via a ping-pong or an ordered reaction mechanism.


Assuntos
Escherichia coli/enzimologia , Sistema Fosfotransferase de Açúcar do Fosfoenolpiruvato/metabolismo , Radioisótopos de Carbono , Cinética , Substâncias Macromoleculares , Manitol/metabolismo , Manitol Fosfatos/metabolismo , Fosfoenolpiruvato/metabolismo , Radioisótopos de Fósforo , Piruvatos/metabolismo , Ácido Pirúvico
18.
Yeast ; 6(3): 245-54, 1990.
Artigo em Inglês | MEDLINE | ID: mdl-2349836

RESUMO

We have cloned the MAS gene, encoding the microbody matrix enzyme malate synthase (EC 4.1.3.2.) from the methylotrophic yeast Hansenula polymorpha. The gene was isolated by screening of a genomic library with a mixed-sequence probe, based on the partial amino acid sequence of the purified enzyme. The nucleotide sequence of a 2.4-kilobase stretch of DNA covering the MAS gene was determined. The gene contains an open reading frame of 555 amino acids, amounting to a calculated molecular mass of 63,254 for the encoded protein. Comparison of the amino acid sequence with the malate synthase sequences of Escherichia coli, Brassica napus L. and Cucumis sativus L. clearly establishes the homology of all four proteins. Compared to the soluble enzyme from E. coli, the malate synthases from H. polymorpha and both plant species, which are located in the microbodies, have a short carboxy-terminal extension. In the plant malate synthases, the extension is probably involved in routing to the microbodies, since it contains the potential peroxisomal targeting signal, Ser-Arg/Lys-Leu, at the carboxy terminus. The H. polymorpha enzyme terminates with similar amino acids, but their sequence, Ser-Leu-Lys, does not conform to any of the known peroxisomal targeting signals.


Assuntos
DNA Fúngico/genética , Malato Sintase/genética , Oxo-Ácido-Liases/genética , Pichia/enzimologia , Saccharomycetales/enzimologia , Sequência de Aminoácidos , Sequência de Bases , Southern Blotting , Clonagem Molecular , Microcorpos/enzimologia , Dados de Sequência Molecular , Pichia/genética , Mapeamento por Restrição
19.
Appl Environ Microbiol ; 67(3): 1292-9, 2001 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-11229924

RESUMO

The marine oligotrophic ultramicrobacterium Sphingomonas alaskensis RB2256 has a physiology that is distinctly different from that of typical copiotrophic marine bacteria, such as Vibrio angustum S14. This includes a high level of inherent stress resistance and the absence of starvation-induced stress resistance to hydrogen peroxide. In addition to periods of starvation in the ocean, slow, nutrient-limited growth is likely to be encountered by oligotrophic bacteria for substantial periods of time. In this study we examined the effects of growth rate on the resistance of S. alaskensis RB2256 to hydrogen peroxide under carbon or nitrogen limitation conditions in nutrient-limited chemostats. Glucose-limited cultures of S. alaskensis RB2256 at a specific growth rate of 0.02 to 0.13 h(-1) exhibited 10,000-fold-greater viability following 60 min of exposure to 25 mM hydrogen peroxide than cells growing at a rate of 0.14 h(-1) or higher. Growth rate control of stress resistance was found to be specific to carbon and energy limitation in this organism. In contrast, V. angustum S14 did not exhibit growth rate-dependent stress resistance. The dramatic switch in stress resistance that was observed under carbon and energy limitation conditions has not been described previously in bacteria and thus may be a characteristic of the oligotrophic ultramicrobacterium. Catalase activity varied marginally and did not correlate with the growth rate, indicating that hydrogen peroxide breakdown was not the primary mechanism of resistance. More than 1,000 spots were resolved on silver-stained protein gels for cultures growing at rates of 0.026, 0.076, and 0.18 h(-1). Twelve protein spots had intensities that varied by more than twofold between growth rates and hence are likely to be important for growth rate-dependent stress resistance. These studies demonstrated the crucial role that nutrient limitation plays in the physiology of S. alaskensis RB2256, especially under oxidative stress conditions.


Assuntos
Peróxido de Hidrogênio/farmacologia , Sphingomonas/efeitos dos fármacos , Sphingomonas/crescimento & desenvolvimento , Catalase/metabolismo , Meios de Cultura/química , Resistência Microbiana a Medicamentos , Eletroforese em Gel Bidimensional , Regulação Bacteriana da Expressão Gênica , Glucose/metabolismo , Água do Mar/microbiologia
20.
Biochemistry ; 22(26): 6163-70, 1983 Dec 20.
Artigo em Inglês | MEDLINE | ID: mdl-6362721

RESUMO

The phosphoryl-group transfer from PHPr to glucose or alpha-methylglucose and from glucose 6-phosphate to these same sugars catalyzed by membrane-bound EIIBGlc of the bacterial phosphoenolpyruvate-dependent phosphotransferase system has been studied in vitro. Kinetic measurements revealed that both the phosphorylation reaction and the exchange reaction proceed according to a ping-pong mechanism in which a phosphorylated membrane-bound enzyme II acts as an obligatory intermediate. The occurrence of a phospho-IIBGlc/IIIGlc has been physically demonstrated by the production of a glucose 6-phosphate burst from membranes phosphorylated by phosphoenolpyruvate, HPr, and EI. The observation of similar second-order rate constants for the production of sugar phosphate starting with different phosphoryl-group donors confirms the catalytic relevance of the phosphoenzyme IIBGlc intermediate. The in vitro results, together with data published by other investigators, have led to a model describing sugar phosphorylation and transport in vivo.


Assuntos
Sistema Fosfotransferase de Açúcar do Fosfoenolpiruvato/metabolismo , Salmonella typhimurium/metabolismo , Bactérias/metabolismo , Transporte Biológico Ativo , Glucose-6-Fosfato , Glucofosfatos/metabolismo , Concentração de Íons de Hidrogênio , Cinética , Modelos Químicos , Fosforilação , Piruvatos/metabolismo , Ácido Pirúvico
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