RESUMO
Stomata in leaves regulate gas (carbon dioxide and water vapor) exchange and water transpiration between plants and the atmosphere. SLow Anion Channel 1 (SLAC1) mediates anion efflux from guard cells and plays a crucial role in controlling stomatal aperture. It serves as a central hub for multiple signaling pathways in response to environmental stimuli, with its activity regulated through phosphorylation via various plant protein kinases. However, the molecular mechanism underlying SLAC1 phosphoactivation has remained elusive. Through a combination of protein sequence analyses, AlphaFold-based modeling and electrophysiological studies, we unveiled that the highly conserved motifs on the N- and C-terminal segments of SLAC1 form a cytosolic regulatory domain (CRD) that interacts with the transmembrane domain(TMD), thereby maintaining the channel in an autoinhibited state. Mutations in these conserved motifs destabilize the CRD, releasing autoinhibition in SLAC1 and enabling its transition into an activated state. Our further studies demonstrated that SLAC1 activation undergoes an autoinhibition-release process and subsequent structural changes in the pore helices. These findings provide mechanistic insights into the activation mechanism of SLAC1 and shed light on understanding how SLAC1 controls stomatal closure in response to environmental stimuli.
Assuntos
Proteínas de Arabidopsis , Arabidopsis , Estômatos de Plantas , Transdução de Sinais , Fosforilação , Estômatos de Plantas/metabolismo , Proteínas de Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Arabidopsis/metabolismo , Arabidopsis/genética , Proteínas de Membrana/metabolismo , Proteínas de Membrana/genética , Domínios Proteicos , MutaçãoRESUMO
The kinetic data of cyclopentadiene C5H6 oxidation reactions are significant for the construction of aromatics oxidation mechanism because cyclopentadiene C5H6 has been proved to be an important intermediate in the aromatics combustion. Kinetics for the elementary reactions on the potential energy surface (PES) relevant for the C5H6 + HO2 reaction are studied in this work. Stationary points on the PES are calculated by employing the CCSD(T)/cc-pVTZ//B3LYP/6-311G(d,p) level of theory. High-pressure limit and pressure-dependent rate constants for elementary reactions on this PES are calculated using conventional transition state theory (TST), variational transition-state theory (VTST) and Rice-Ramsberger-Kassel-Marcus/master equation (RRKM/ME) theory. In this work, the reaction channels for the C5H6 + HO2 reaction, which include H-abstraction channels from C5H6 by HO2 to form the C5H5 + H2O2 and the addition channels through well-skipping pathways to form the bimolecular products C5H7 + O2 or C5H6O + OH, or through C5H7O2 stabilization and its unimolecular decomposition to form the bimolecular products C5H7 + O2 or C5H6O + OH, namely sequential pathways, are studied. Also, the consuming reaction channels for the compounds C5H6O and C5H7 in the addition products are studied. The dominant reaction channels for these reactions are unraveled through comparing the energy barriers and rate constants of all elementary reactions and it is found: (1) HO2 addition to cyclopentadiene C5H6 is more important than direct H-abstraction. (2) in the HO2 addition channels, the well-skipping pathways and sequential pathways are competing and the well-skipping pathways will be favor in the higher pressures and the sequential pathways will be favor in the higher temperature. (3) The major consumption reaction channel for the five-member-ring compound C5H6O is the reaction channel to form C4H6 + CO and the major consumption reaction channel for the five-member-ring compound C5H7 is the reaction channel to form C3H5 + C2H2. High-pressure limit rate constants and pressure-dependent rate constants for elementary reactions on the PES are calculated, which will be useful in modeling the oxidation of aromatic compounds at low- and medium-temperatures.
RESUMO
In this paper, single molecular junctions of Para-phthalic acid and Meta-phthalic acid with Au electrodes were studied by STM break junction approach. Conductance values of 10-3.55 G0 and 10-3.70 G0 were found for Para-phthalic acid and Meta-phthalic acid, respectively. The conductance order between Para-phthalic acid and Meta-phthalic acid with Au is different from that with Cu, which can be contributed to the different coupling between molecules and electrodes; different through-space interaction is proposed for such phenomenon between Cu and Au electrodes. Furthermore, the breaking off distances can reflect the length of molecules. The current work presents the important role of electrode in single molecular junctions with different position anchoring groups.
RESUMO
In this study, the feasibility of applying a magnetotactic bacterial isolate (MTB), Stenotrophomonas sp. to the removal of Au(III) was investigated. Biosorption experiments showed that Au(III) biosorption capacity exhibited no significant difference in the initial pH range of 1.0-5.5, while decreased more significantly in the initial pH range of 5.5-13.0. Langmuir isotherm indicated that the maximum Au(III) biosorption capacity of Stenotrophomonas sp. were 506, 369 and 308 mg g(-1) dry weight biomass at the initial pH values of 2.0, 7.0 and 12.0, respectively. Thiourea was proved to be an effective desorbent to recover Au from the MTB biomass and 91% Au adsorbed on the biomass could be recovered at equilibrium when the thiourea concentration was 0.8M. The magnetic separator developed by our research team used for separating Au loaded MTB biomass showed high separation efficiency, with 100% biomass removed at the magnetic intensity of 1200 Gs in 180 min. The analyses from FTIR and XRD further confirmed that the reduction of Au(III) to Au(0) by the reductants on the MTB biomass occurred, and the deposition of nano-crystal Au(0) particles, ranging from 24.7 to 31.4 nm, could be estimated on the biomass surface.
Assuntos
Poluentes Ambientais/isolamento & purificação , Poluentes Ambientais/metabolismo , Ouro/isolamento & purificação , Ouro/metabolismo , Magnetismo , Esgotos , Stenotrophomonas/metabolismo , Absorção , Biodegradação Ambiental , Biomassa , Mecânica , Reprodutibilidade dos Testes , Água/químicaRESUMO
The aim of the present study was to investigate the optimal mobilization plan in autologous peripheral blood stem cell transplantation for the treatment of diabetic foot and to observe its clinical curative effect. A total of 127 patients with diabetic foot were treated with different doses of granulocyte colony stimulating factor (G-CSF) to mobilize their hematopoietic stem cells. Subsequently, the extracted stem cell suspension was injected into the ischemic lower extremities along the blood vessels in the areas presenting with pathological changes. Following the treatment, the intermittent claudication distance, skin temperature, ankle brachial index and pain scores of the patients were evaluated. In addition, the associations among the mobilization time, doses and peripheral blood CD34+ level were analyzed. The collection efficiency of the stem cells was associated with the dose of G-CSF and the mobilization time. Following the injection of the autologous peripheral blood stem cell suspension, the ischemic area of the patients was improved significantly. In conclusion, autologous peripheral blood stem cell transplantation can promote the establishment of collateral circulation in patients with diabetic foot, and the optimal time for gathering stem cells is closely correlated with the peripheral blood CD34+ level.
RESUMO
Trichloroethylene (TCE), a common groundwater pollutant, was cometabolized by microorganisms in the presence of toluene as a growth substrate. The effect of concentrations of toluene and TCE and temperature on biodegradation was discussed. Acclimated microorganisms degraded TCE after a lag period of 5 to 22 h depending on toluene concentrations. Approximately 60%, 90% and 64% of TCE were degraded at toluene to TCE concentration ratios of 23:1, 115:1 and 230:1, respectively. At a TCE concentration of 1.46 microg/ml, 80% of TCE and 98.4% of toluene were removed. But less degradation of TCE and toluene was observed when TCE concentration was above 48.8 microg/ml. The lag time of TCE decreased and the TCE biodegradation rates increased with the increase of temperature.
Assuntos
Poluentes do Solo/metabolismo , Solventes/metabolismo , Tolueno/química , Tricloroetileno/metabolismo , Poluentes Químicos da Água/metabolismo , TemperaturaRESUMO
The effects of initial glucose concentration and calcium lactate concentration on the lactic acid production by the parent strain, Lactobacillus lactis BME5-18, were studied. The results of the experiments indicated that glucose and lactate repressed the cell growth and the lactic acid production by Lactobacillus lactis BME5-18. A L(+)-lactic acid overproducing strain, Lactobacillus lactis BME5-18M, was screened by mutagenizing the parent strain with ultraviolet (UV) light irradiation and selecting the high glucose and lactate calcium concentration repression resistant mutant. Starting with a concentration of 100g L(-1) glucose, the mutant produced 98.6 g L(-1) lactic acid after 60 h in flasks, 73.9% higher than that of the parent strain. The L(+)-lactic acid purity was 98.1% by weight based on the amount of total lactic acid. The culture of the parent strain could not be analyzed well by conventional metabolic flux analysis techniques, since some pyruvate were accumulated intracellularly. Therefore, a revised flux analysis method was proposed by introducing intracellular pyruvate pool. Further studies demonstrate that there is a high level of NADH oxidase activity (12.11 mmol mg(-1) min(-1)) in the parent strain. The molecular mechanisms of the strain improvement were proposed, i.e., the high level of NADH oxidase activity was eliminated and the uptake rate of glucose was increased from 82.1 C-mmol (g DW h)(-1) to 98.9 C-mmol (g DW h)(-1) by mutagenizing the parent strain with UV, and therefore the mutant strain converts mostly pyruvate to lactic acid with a higher productivity (1.76 g L(-1) h(-1)) than the parent strain (0.95 g L(-1) h(-1)).
Assuntos
Metabolismo Energético/fisiologia , Melhoramento Genético/métodos , Ácido Láctico/biossíntese , Lactobacillus/genética , Lactobacillus/metabolismo , Modelos Biológicos , Complexos Multienzimáticos/metabolismo , NADH NADPH Oxirredutases/metabolismo , Simulação por Computador , Regulação Bacteriana da Expressão Gênica/fisiologia , Regulação Bacteriana da Expressão Gênica/efeitos da radiação , Glucose/metabolismo , Lactobacillus/efeitos da radiação , Mutagênese/efeitos da radiação , Transdução de Sinais/fisiologia , Transdução de Sinais/efeitos da radiação , Especificidade da Espécie , Raios UltravioletaRESUMO
A fed-batch fermentation of Lactobacillus lactis to produce L-lactic acid was developed in which the residual glucose concentration in the culture was used to control a continuous feeding strategy. Up to 210 g L-lactic acid l(-1) (97% yield) was obtained. The maximal dry cell was 2.7 g l(-1) and the average L-lactic acid productivity was 2 g l(-1) h(-1).