Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 6 de 6
Filtrar
Mais filtros

Base de dados
Tipo de documento
Intervalo de ano de publicação
1.
Biochim Biophys Acta ; 1847(2): 276-285, 2015 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-25481108

RESUMO

Cytb559 in Photosystem II is a heterodimeric b-type cytochrome. The subunits, PsbE and PsbF, consist each in a membrane α-helix. Roles for Cytb559 remain elusive. In Thermosynechococcus elongatus, taking advantage of the robustness of the PSII variant with PsbA3 as the D1 subunit (WT*3), 4 mutants were designed hoping to get mutants nevertheless the obligatory phototrophy of this cyanobacterium. In two of them, an axial histidine ligand of the haem-iron was substituted for either a methionine, PsbE/H23M, which could be potentially a ligand or for an alanine, PsbE/H23A, which cannot. In the other mutants, PsbE/Y19F and PsbE/T26P, the environment around PsbE/H23 was expected to be modified. From EPR, MALDI-TOF and O2 evolution activity measurements, the following results were obtained: Whereas the PsbE/H23M and PsbE/H23A mutants assemble only an apo-Cytb559 the steady-state level of active PSII was comparable to that in WT*3. The lack of the haem or, in PsbE/T26P, conversion of the high-potential into a lower potential form, slowed-down the recovery rate of the O2 activity after high-light illumination but did not affect the photoinhibition rate. This resulted in the following order for the steady-state level of active PSII centers under high-light conditions: PsbE/H23M≈PsbE/H23A<< PsbE/Y19F≤PsbE/T26P≤WT*3. These data show i) that the haem has no structural role provided that PsbE and PsbF are present, ii) a lack of correlation between the rate of photoinhibition and the Em of the haem and iii) that the holo-Cytb559 favors the recovery of a functional enzyme upon photoinhibition.


Assuntos
Grupo dos Citocromos b/química , Complexo de Proteína do Fotossistema II/química , Sequência de Aminoácidos , Sequência de Bases , Grupo dos Citocromos b/fisiologia , Espectroscopia de Ressonância de Spin Eletrônica , Luz , Dados de Sequência Molecular , Oxigênio/metabolismo , Complexo de Proteína do Fotossistema II/fisiologia
2.
Biochim Biophys Acta ; 1837(1): 139-48, 2014 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-24060528

RESUMO

In Photosystem II (PSII) of the cyanobacterium Thermosynechococcus elongatus, glutamate 130 in the high-light variant of the D1-subunit (PsbA3) was changed to glutamine in a strain lacking the two other genes for D1, psbA1 and psbA2. The resulting PSII (PsbA3/Glu130Gln) was compared with those from the "native" high-light (PsbA3-PSII) and low-light (PsbA1-PSII) variants, which differ by 21 amino acid including Glu130Gln. H-bonding from D1-Glu130Gln to the primary electron acceptor, PheophytinD1 (PheoD1), is known to affect the Em of the PheoD1/PheoD1(-) couple. The Gln130 mutation here had little effect on water splitting, charge accumulation and photosensitivity but did slow down S2QA(-) charge recombination and up-shift the thermoluminescence while increasing its yield. These changes were consistent with a ≈-30mV shift of the PheoD1/PheoD1(-)Em, similar to earlier single site-mutation results from other species and double the ≈-17mV shift seen for PsbA1-PSII versus PsbA3-PSII. This is attributed to the influence of the other 20 amino-acids that differ in PsbA3. A computational model for simulating S2QA(-) recombination matched the experimental trend: the S2QA(-) recombination rate in PsbA1-PSII differed only slightly from that in PsbA3-PSII, while in Glu130-PsbA3-PSII there was a more pronounced slowdown of the radical pair decay. The simulation predicted a major effect of the PheoD1/PheoD1(-) potential on (1)O2 yield (~60% in PsbA1-PSII, ~20% in PsbA3-PSII and ~7% in Gln130-PsbA3-PSII), reflecting differential sensitivities to high light.


Assuntos
Cianobactérias/química , Feofitinas/química , Complexo de Proteína do Fotossistema II/química , Cianobactérias/metabolismo , Transporte de Elétrons , Ácido Glutâmico/genética , Glutamina/genética , Luz , Mutação , Oxirredução , Feofitinas/metabolismo , Complexo de Proteína do Fotossistema II/genética , Complexo de Proteína do Fotossistema II/metabolismo
3.
Biochim Biophys Acta ; 1827(10): 1174-82, 2013 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-23770319

RESUMO

Cyanobacteria have multiple psbA genes encoding PsbA, the D1 reaction center protein of the Photosystem II (PSII) complex. The thermophilic cyanobacterium Thermosynechococcus elongatus has three psbA genes differently expressed depending on the environmental conditions. Among the 344 residues of PsbA, there are 21 substitutions between PsbA1 and PsbA3, 31 between PsbA1 and PsbA2 and 27 between PsbA2 and PsbA3. In this study, we found a new hemoprotein that is expressed when the T. elongatus genome has only the psbA2 gene for D1. This hemoprotein was found in both the non-membrane proteins and associated to the purified PsbA2-PSII core complex. This protein could be removed by the washing of PSII with Tris-washing or CaCl2-washing. From MALDI-TOF/TOF spectrometry, N-terminal sequencing and MALDI-MS/MS analysis upon tryptic digestion, the new hemoprotein was identified to be the tll0287 gene product with a molecular mass close to 19kDa. Until now, tll0287 was registered as a gene encoding a hypothetical protein with an unknown function. From the amino acid sequence and the EPR spectrum the 5th and 6th axial ligands of the heme iron are the His145 and likely either the Tyr93, Tyr159 or Tyr165, respectively. From EPR, the heme containing Tll0287 protein associated to PsbA2-PSII corresponds to approximately 25% of the Cytc550 content whereas, from SDS page analysis, the total amount of Tll0287 with and without the heme seems almost in a stoichiometric amount with PsbA2-PSII. Homologous genes to tll0287 are found in several cyanobacteria. Possible roles for Tll0287 are suggested.


Assuntos
Hemeproteínas/metabolismo , Complexo de Proteína do Fotossistema II/metabolismo , Synechococcus/metabolismo , Sequência de Aminoácidos , Cromatografia em Gel , Espectroscopia de Ressonância de Spin Eletrônica , Transporte de Elétrons , Dados de Sequência Molecular , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz
4.
Biochim Biophys Acta Bioenerg ; 1865(1): 149013, 2024 01 01.
Artigo em Inglês | MEDLINE | ID: mdl-37717932

RESUMO

Photosystem II is the water/plastoquinone photo-oxidoreductase of photosynthesis. The photochemistry and catalysis occur in a quasi-symmetrical heterodimer, D1D2, that evolved from a homodimeric ancestor. Here, we studied site-directed mutants in PSII from the thermophilic cyanobacterium Thermosynechoccocus elongatus, focusing on the primary electron donor chlorophyll a in D1, ChlD1, and on its symmetrical counterpart in D2, ChlD2, which does not play a direct photochemical role. The main conserved amino acid specific to ChlD1 is D1/T179, which H-bonds the water ligand to its Mg2+, while its counterpart near ChlD2 is the non-H-bonding D2/I178. The symmetrical-swapped mutants, D1/T179I and D2/I178T, and a second ChlD2 mutant, D2/I178H, were studied. The D1 mutations affected the 686 nm absorption attributed to ChlD1, while the D2 mutations affected a 663 nm feature, tentatively attributed to ChlD2. The mutations had little effect on enzyme activity and forward electron transfer, reflecting the robustness of the overall enzyme function. In contrast, the mutations significantly affected photodamage and protective mechanisms, reflecting the importance of redox tuning in these processes. In D1/T179I, the radical pair recombination triplet on ChlD1 was shared onto a pheophytin, presumably PheD1 and the detection of 3PheD1 supports the proposed mechanism for the anomalously short lifetime of 3ChlD1; e.g. electron transfer quenching by QA- of 3PheD1 after triplet transfer from 3ChlD1. In D2/I178T, a charge separation could occur between ChlD2 and PheD2, a reaction that is thought to occur in ancestral precursors of PSII. These mutants help understand the evolution of asymmetry in PSII.


Assuntos
Aminoácidos , Complexo de Proteína do Fotossistema II , Complexo de Proteína do Fotossistema II/metabolismo , Aminoácidos/genética , Clorofila A , Clorofila/metabolismo , Mutagênese Sítio-Dirigida , Água
5.
Respir Investig ; 62(6): 1058-1063, 2024 Sep 20.
Artigo em Inglês | MEDLINE | ID: mdl-39305533

RESUMO

BACKGROUND: Nontuberculous mycobacterial pulmonary disease (NTM-PD) patients often have exercise intolerance. Pulmonary rehabilitation (PR) to improve such patients' conditions is often not based on its exercise pathophysiology. We have reported that the oxygen consumption (ΔFO2) by expiratory gas analysis, i.e., the inspired-expired-expiratory mean oxygen concentration difference, is related to the minute ventilation-carbon dioxide output (V'E-V'CO2)-slope and oxygen uptake (V'O2) independent of the V'E. The aim of this study was to investigate how ΔFO2 is related to dynamic ventilatory variables, chest computed tomography (CT), and echocardiography findings in NTM-PD patients to understand their pathophysiological conditions. METHODS: Clinical data of NTM-PD patients with exertional dyspnea (n = 29) who underwent incremental exercise testing, chest CT, and echocardiography at the same time were compared with those of control participants (n = 12). RESULTS: In the NTM-PD group, 1) peak V'O2 decreased (NTM-PD: 17.6 vs. controls: 28.7 mL⋅min-1⋅kg-1), and 2) ΔFO2 at peak exercise was negatively correlated with respiratory frequency at peak exercise (correlation coefficient: r = -0.80, p < 0.0001), V'E-V'CO2-slope (r = -0.75, p < 0.0001), bronchiectasis CT score (r = -0.52, p = 0.0042), and the trans-tricuspid pressure gradient (r = -0.39, p = 0.0417), and positively correlated with peak V'O2 (r = 0.71, p < 0.0001) and the body mass index (r = 0.42, p = 0.0217), but it was not correlated with V'E at peak exercise and the cavity CT score. CONCLUSIONS: Exertional oxygen consumption, independent of ventilatory ability, is associated with exercise tolerance and ventilatory efficiency, while being related to tachypnea and bronchiectasis rather than cavitation in NTM-PD patients. These findings may be useful in considering exercise physiology-based PR for NTM-PD patients with exertional dyspnea.

6.
Biochemistry ; 52(52): 9426-31, 2013 Dec 31.
Artigo em Inglês | MEDLINE | ID: mdl-24320870

RESUMO

The electron density map of the 3D crystal of Photosystem II from Thermosynechococcus vulcanus with a 1.9 Šresolution (PDB: 3ARC ) exhibits, in the two monomers in the asymmetric unit cell, an, until now, unidentified and uninterpreted strong difference in electron density centered at a distance of around 1.5 Šfrom the nitrogen Nδ of the imidazole ring of D2-His336. By MALDI-TOF/MS upon tryptic digestion, it is shown that ~20-30% of the fragments containing the D2-His336 residue of Photosystem II from both Thermosynechococcus vulcanus and Thermosynechococcus elongatus bear an extra mass of +16 Da. Such an extra mass likely corresponds to an unprecedented post-translational or chemical hydroxyl modification of histidine.


Assuntos
Proteínas de Bactérias/metabolismo , Cianobactérias/metabolismo , Histidina/metabolismo , Radical Hidroxila/metabolismo , Complexo de Proteína do Fotossistema II/metabolismo , Motivos de Aminoácidos , Proteínas de Bactérias/química , Proteínas de Bactérias/genética , Cristalografia por Raios X , Cianobactérias/química , Cianobactérias/genética , Histidina/química , Histidina/genética , Modelos Moleculares , Peso Molecular , Complexo de Proteína do Fotossistema II/química , Complexo de Proteína do Fotossistema II/genética
SELEÇÃO DE REFERÊNCIAS
Detalhe da pesquisa