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Simultaneous CO2 sequestration and nitrate removal can be achieved by co-cultivation of Chlorella vulgaris with Pseudomonas sp. However, a comprehensive understanding of the synergistic mechanism between C. vulgaris and Pseudomonas sp. remains unknown. In this study, transcriptomics and metabolomics analysis were employed to elucidate the synergistic mechanism of C. vulgaris and Pseudomonas sp. Transcriptomic and metabolomic analyses identified 3664 differentially expressed genes and 314 metabolites. Transcriptome analysis revealed that co-culture with Pseudomonas sp. promoted the photosynthesis of C. vulgaris by promoting the synthesis of photosynthetic pigments and photosynthesis-antenna proteins. Furthermore, it stimulated pathways associated with energy metabolism from carbon sources, such as the Calvin cycle, glycolytic pathway, and TCA cycle. Additionally, Pseudomonas sp. reduced nitrate levels in the co-culture system by denitrification, and microalgae regulated nitrate uptake by down-regulating the transcript levels of nitrate transporter genes. Metabolomic analysis indicated that nutrient exchange was conducted between algae and bacteria, and amino acids, phytohormones, and organic heterocyclic compounds secreted by the bacteria promoted the growth metabolism of microalgae. After supplementation with differential metabolites, the carbon fixation rate and nitrate removal rate of the co-culture system reached 0.549 g L-1 d-1 and 135.4 mg L-1 d-1, which were increased by 20% and 8%, respectively. This study provides a theoretical insight into microalgae-bacteria interaction and its practical application, as well as a novel perspective on flue gas treatment management.
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Microalgae are considered to be natural producers of bioactive pigments, with the production of pigments from microalgae being a sustainable and economical strategy that promises to alleviate growing demand. Chlorophyll, as the main pigment of photosynthesis, has been widely studied, but its medicinal applications as an antioxidant, antibacterial, and antitumor reagent are still poorly understood. Chlorophyll is the most important pigment in plants and algae, which not only provides food for organisms throughout the biosphere, but also plays an important role in a variety of human and man-made applications. The biological activity of chlorophyll is closely related to its chemical structure; its specific structure offers the possibility for its medicinal applications. This paper reviews the structural and functional roles of microalgal chlorophylls, commonly used extraction methods, and recent advances in medicine, to provide a theoretical basis for the standardization and commercial production and application of chlorophylls.
Assuntos
Microalgas , Humanos , Clorofila/química , Fotossíntese , Antioxidantes/farmacologia , Antioxidantes/química , PlantasRESUMO
Carbon dioxide and nitrogen oxides are the main components of fossil flue gas causing the most serious environmental problems. Developing a sustainable and green method to treat carbon dioxide and nitrogen oxides of flue gas is still challenging. Here, a co-cultured microalgae/bacteria system, Chlorella vulgaris and Pseudomonas sp., was developed for simultaneous sequestration of CO2 and removal of nitrogen oxides from flue gas, as well as producing valuable microalgae biomass. The co-cultured Chlorella vulgaris and Pseudomonas sp. showed the highest CO2 fixation and NO3--N removal rate of 0.482 g L-1d-1 and 129.6 mg L-1d-1, the total chlorophyll accumulation rate of 65.6 mg L-1 at the initial volume ratio of Chlorella vulgaris and Pseudomonas sp. as 1:10. The NO3--N removal rate can be increased to 183.5 mg L-1d-1 by continuous addition of 0.6 g L-1d-1 of glucose, which was 37% higher than that of co-culture system without the addition of glucose. Photosynthetic activity and carbonic anhydrase activity of Chlorella vulgaris were significantly increased when co-cultured with Pseudomonas sp. Excitation-emission matrix (EEM) fluorescence spectroscopy indicated that the humic acid-like substances released from Pseudomonas sp. could increase the growth of microalgae. This work provides an attractive way to simultaneously treatment of CO2 and NOX from flue gas to produce valuable microalgal biomass.
Assuntos
Chlorella vulgaris , Microalgas , Dióxido de Carbono , Nitratos , Óxidos de Nitrogênio , Técnicas de Cocultura , Biomassa , Sequestro de CarbonoRESUMO
Oligomeric organization of G protein-coupled receptors is proposed to regulate receptor signaling and function, yet rapid and precise identification of the oligomeric status especially for native receptors on a cell membrane remains an outstanding challenge. By using blinking carbon dots (CDs), we now develop a deep learning (DL)-based blinking fingerprint recognition method, named deep-blinking fingerprint recognition (BFR), which allows automatic classification of CD-labeled receptor organizations on a cell membrane. This DL model integrates convolutional layers, long-short-term memory, and fully connected layers to extract time-dependent blinking features of CDs and is trained to a high accuracy (â¼95%) for identifying receptor organizations. Using deep blinking fingerprint recognition, we found that CXCR4 mainly exists as 87.3% monomers, 12.4% dimers, and <1% higher-order oligomers on a HeLa cell membrane. We further demonstrate that the heterogeneous organizations can be regulated by various stimuli at different degrees. The receptor-binding ligands, agonist SDF-1α and antagonist AMD3100, can induce the dimerization of CXCR4 to 33.1 and 20.3%, respectively. In addition, cytochalasin D, which inhibits actin polymerization, similarly prompts significant dimerization of CXCR4 to 30.9%. The multi-pathway organization regulation will provide an insight for understanding the oligomerization mechanism of CXCR4 as well as for elucidating their physiological functions.
Assuntos
Carbono , Aprendizado Profundo , Pontos Quânticos , Receptores CXCR4 , Benzilaminas/química , Benzilaminas/farmacologia , Quimiocina CXCL12/agonistas , Ciclamos/química , Ciclamos/farmacologia , Células HeLa , Humanos , Receptores CXCR4/químicaRESUMO
BACKGROUND: Due to its high expression capability, recombination of Escherichia coli and pET vector has become the bioengineering preferred expression system. Because ß-lactamases mediate bacterial antimicrobial resistance, these enzymes have a substantial clinical impact. Using the E. coli expression system, several kinds of ß-lactamases have been produced. However, previous studies have been focused on characterizing target ß-lactamases, and the effects of cultivation and induction conditions on the expression efficiency of target enzymes were not addressed. RESULTS: Using pET-28a as the cloning vector and E. coli BL21(DE3) as the expression host, this study originally elucidated the effects of IPTG concentration, culture temperature, induction time, and restriction sites on recombinant ß-lactamase expression. Moreover, the effects of the target protein length and the 6 × His-tag fusion position on enzyme purification were also explored, and consequently, this study yielded several important findings. (i) Only the signal peptide-detached recombinant ß-lactamase could exist in a soluble form. (ii) Low-temperature induction was beneficial for soluble ß-lactamase expression. (iii) The closer to the rbs the selected restriction site was, the more difficult it was to express soluble ß-lactamase. (iv) The short-chain recombinant protein and the protein with His-tag fused at its C-terminus showed high affinity to the Ni2+ column. CONCLUSIONS: Based on our findings, researchers can easily design an effective program for the high production of soluble recombinant ß-lactamases to facilitate other related studies.
Assuntos
Escherichia coli , beta-Lactamases , beta-Lactamases/genética , Escherichia coli/genética , Antibacterianos , Bioengenharia , Vetores Genéticos/genéticaRESUMO
Single-molecule fluorescence imaging is a powerful tool to study protein function by tracking molecular position and distribution, but the precise and rapid identification of dynamic molecules remains challenging due to the heterogeneous distribution and interaction of proteins on the live cell membrane. We now develop a deep-learning (DL)-assisted single-molecule imaging method that can precisely distinguish the monomer and complex for rapid and real-time tracking of protein interaction. This DL-based model, which comprises convolutional layers, max pooling layers, and fully connected layers, is trained to reach an accuracy of >98% for identifying monomer and complex. We use this method to investigate the dynamic process of chemokine receptor CXCR4 on the live cell membrane during the early signaling stage. The results show that, upon ligand activation, the CXCR4 undergoes a dynamic process of forming a receptor complex. We further demonstrate that the CXCR4 complex tends to be internalized at 2.5-fold higher rate into the cell interior than the monomer via the clathrin-dependent pathway. This study is the first example to scrutinize the early signaling process of CXCR4 at the single-molecule level on the live cell membrane. We envision that this DL-assisted imaging method would be a broadly useful technique to study more protein families for elucidating their physiological and pathological functions.
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Collagen plays an important role in the formation of extracellular matrix (ECM) and development/migration of cells and tissues. Here we report the preparation of collagen and collagen hydrogel from the skin of tilapia and an evaluation of their potential as a wound dressing for the treatment of refractory wounds. The acid-soluble collagen (ASC) and pepsin-soluble collagen (PSC) were extracted and characterized using sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE), differential scanning calorimetry (DSC), circular dichroism (CD) and Fourier transform infrared spectroscopy (FTIR) analysis. Both ASC and PSC belong to type I collagen and have a complete triple helix structure, but PSC shows lower molecular weight and thermal stability, and has the inherent low antigenicity. Therefore, PSC was selected to prepare biomedical hydrogels using its self-aggregating properties. Rheological characterization showed that the mechanical strength of the hydrogels increased as the PSC content increased. Scanning electron microscope (SEM) analysis indicated that hydrogels could form a regular network structure at a suitable PSC content. Cytotoxicity experiments confirmed that hydrogels with different PSC content showed no significant toxicity to fibroblasts. Skin repair experiments and pathological analysis showed that the collagen hydrogels wound dressing could significantly accelerate the healing of deep second-degree burn wounds and the generation of new skin appendages, which can be used for treatment of various refractory wounds.
Assuntos
Bandagens , Queimaduras/terapia , Ciclídeos , Colágeno Tipo I/farmacologia , Proteínas de Peixes/farmacologia , Animais , Colágeno Tipo I/isolamento & purificação , Colágeno Tipo I/ultraestrutura , Modelos Animais de Doenças , Feminino , Proteínas de Peixes/isolamento & purificação , Proteínas de Peixes/ultraestrutura , Humanos , Hidrogéis/farmacologia , Masculino , Microscopia Eletrônica de Varredura , Ratos , Pele/química , Pele/lesões , Cicatrização/efeitos dos fármacosRESUMO
To regulate the effectiveness of photosynthesis and photoprotection cyanobacteria utilize a system consisting of only few components. Photoactivation of the orange carotenoid protein (OCP) enables its interaction with a specific, yet controversial site in the core of the light-harvesting antenna, the phycobilisome (PBS). The resulting delivery of a quenching carotenoid molecule to the antenna pigments leads to thermal dissipation of the excitation energy absorbed by the latter, and, consequently, to depression of the photosynthetic activity. The nature of the OCP-induced PBS fluorescence quenching mechanism remains debatable, however, specific protein-protein interactions between PBS and photoactivated OCP should provide a unique environment for interactions between the excitation energy donor and acceptor. Here we questioned whether the Förster theory of resonance energy transfer can explain PBS quenching by OCP even at their very small spectral overlap and whether in model systems, the absence of specific protein-protein interactions of OCP with a donor of energy can be compensated by a better spectral overlap. Hybridization of algal R-phycoerythrin with cyanobacterial OCP by chemical crosslinking results in a significant decrease of R-phycoerythrin fluorescence lifetime, irrespective of the OCP photoactivation status. Supported by structural considerations, this indicates that FRET may be the essence of cyanobacterial photoprotection mechanism.
Assuntos
Proteínas de Algas/metabolismo , Proteínas de Bactérias/metabolismo , Transferência Ressonante de Energia de Fluorescência/métodos , Ficoeritrina/metabolismo , Proteínas de Algas/química , Proteínas de Bactérias/química , Carotenoides/química , Carotenoides/metabolismo , Luz , Fotossíntese/efeitos da radiação , Ficobilissomas/química , Ficobilissomas/metabolismo , Ficoeritrina/química , Porphyra/química , Espectrometria de Fluorescência/métodosRESUMO
A growing number of proteins have been identified as knotted in their native structures, with such entangled topological features being expected to play stabilizing roles maintaining both the global fold and the nature of proteins. However, the molecular mechanism underlying the stabilizing effect is ambiguous. Here, we combine unbiased and mechanical atomistic molecular dynamics simulations to investigate how a protein is stabilized by an inherent knot by directly comparing chemical, thermal, and mechanical denaturing properties of two proteins having the same sequence and secondary structures but differing in the presence or absence of an inherent knot. One protein is YbeA from Escherichia coli, containing a deep trefoil knot within the sequence, and the other is the modified protein with the knot of YbeA being removed. Under certain chemical denaturing conditions, the unknotted protein fully unfolds whereas the knotted protein does not, suggesting a higher intrinsic stability for the protein having a knot. Both proteins unfold under enhanced thermal fluctuations but at different rates and with distinct pathways. Opening the hydrophobic core via separation between two α-helices is identified as a crucial step initiating the protein unfolding, which, however, is restrained for the knotted protein by topological and geometrical frustrations. Energy barriers for denaturing the protein are reduced by removing the knot, as evidenced by mechanical unfolding simulations. Finally, yet importantly, no obvious change in size or location of the knot was observed during denaturing processes, indicating that YbeA may remain knotted for a relatively long time during and after denaturation.
Assuntos
Proteínas de Escherichia coli/química , Metiltransferases/química , Simulação de Dinâmica Molecular , Fenômenos Biomecânicos , Desnaturação Proteica , Estabilidade Proteica , Estrutura Secundária de Proteína , TemperaturaRESUMO
The relationship between the oligomeric status and functions of chemokine receptor CCR3 is still controversial. We use total internal reflection fluorescence microscopy at the single-molecule level to visualize the oligomeric status of CCR3 and its regulation of the membrane of stably transfected T-REx-293 cells. We find that the population of the dimers and oligomers of CCR3 can be modulated by the binding of ligands. Natural agonists can induce an increase in the level of dimers and oligomers at high concentrations, whereas antagonists do not have a significant influence on the oligomeric status. Moreover, monomeric CCR3 exhibits a stronger chemotactic response in the migration assay of stably transfected CCR3 cells. Together, these data support the notion that CCR3 exists as a mixture of monomers and dimers under nearly physiological conditions and the monomeric CCR3 receptor is the minimal functional unit in cellular signaling transduction. To the best of our knowledge, these results constitute the first report of the oligomeric status of CCR3 and its regulation.
Assuntos
Microscopia de Fluorescência/métodos , Receptores CCR3/metabolismo , Imagem Individual de Molécula/métodos , Linhagem Celular , Membrana Celular/metabolismo , Quimiotaxia , Relação Dose-Resposta a Droga , Humanos , Ligantes , Fotodegradação , Multimerização Proteica , Receptores CCR3/agonistas , Receptores CCR3/antagonistas & inibidores , Proteínas Recombinantes/metabolismo , Transdução de SinaisRESUMO
Nanotechnology-enhanced drug delivery via receptor-mediated endocytosis provides a promising and clinically translatable strategy to targeted diagnosis and precise therapy, yet an in-depth understanding of this process is technically limited by our inability to probe the nanocarrier distributions at the cell surface and inside the cell at nanoscale resolution. Here, we report small blinking single-layer graphene oxide nanosheet (GONS) that serves both as a nanoscopy fluorophore and as a drug-bearing nanocarrier for addressing such a task. The GONS blinks spontaneously with a low duty cycle (â¼0.003), high photon output (â¼3000 photons per switching event), and higher photostability than organic dyes, thus affording well for single molecule localization-based super-resolution imaging. Applying the localization analysis, we reveal GONS clustering size, GONS number in each cluster, and the number fraction of GONSs that participate in clustering at the cell surface and in the cytoplasm, respectively, and track their evolutions over 24 h. The data suggest that the nanocarrier clustering and distribution at the cell surface control their endocytosis and accumulation inside the cell. This process is drug-independent during which drug transportation into the destination relies on its own loading and escaping capability. Thus, this work demonstrates the great potential of the dual-functional GONS in quantitative super-resolution imaging of drug carriers in cells, which is helpful for the rational design of a smart drug delivery system aiming at achieving full therapeutic capacity.
Assuntos
Portadores de Fármacos , Grafite/administração & dosagem , Nanopartículas , Linhagem Celular Tumoral , Endocitose , Corantes Fluorescentes/química , Células HeLa , HumanosRESUMO
A bacterial strain, K11T, capable of degrading phenol derivatives was isolated from activated sludge of a sewage treatment plant in China. This strain, which can degrade more than ten phenol derivatives, was identified as a Gram-stain negative, rod-shaped, asporogenous, facultative anaerobic bacterium with a polar flagellum. The strain was found to grow in tryptic soy broth in the presence of 0-2.5% (w/v) NaCl (optimum 0-1%), at 4-43 °C (optimum 30-35 °C) and pH 4.5-10.5 (optimum 7.5-8). Comparative analysis of nearly full-length 16S rRNA gene sequences showed that this strain belongs to the genus Thauera. The 16S rRNA gene sequence was found to show high similarity (97.5%) to that of Thauera chlorobenzoica 3CB-1T, with lesser similarity to other recognised Thauera strains. The G+C content of the DNA of the strain was determined to be 67.8 mol%. The DNA-DNA hybridization value between K11T and Thauera aromatica DSM6984T was 10.4 ± 4.5%. The genomic OrthoANI values of K11T with the other nine type strains of genus Thauera were less than 81.1%. Chemotaxonomic analysis of strain K11T revealed that Q-8 is the predominant quinone; the polar lipids contain phosphatidylglycerol, diphosphatidylglycerol, phosphatidylethanolamine, two unidentified phospholipids and five uncharacterised lipids; the major cellular fatty acid was identified as summed feature 3 (C16:1 ω7c and/or iso-C15:0 2-OH; 45.9%), followed by C16:0 (20.5%) and C18:1 ω7c (15.8%). Based on the phenotypic and phylogenetic evidence, DNA-DNA hybridisation, OrthoANI, chemotaxonomic analysis and results of the physiological and biochemical tests, a new species named Thauera sinica sp. nov. is proposed with strain K11T (= CGMCC 1.15731T = KACC 19216T) designated as the type strain.
Assuntos
Thauera/genética , Técnicas de Tipagem Bacteriana , Composição de Bases/genética , DNA Bacteriano/genética , RNA Ribossômico 16S/genética , Esgotos/microbiologia , Thauera/isolamento & purificaçãoRESUMO
BACKGROUND: Chemokines and their cognate receptors play important role in the control of leukocyte chemotaxis, HIV entry and other inflammatory diseases. Developing an effcient method to investigate the functional expression of chemokines and its interactions with specific receptors will be helpful to asses the structural and functional characteristics as well as the design of new approach to therapeutic intervention. RESULTS: By making systematic optimization study of expression conditions, soluble and functional production of chemokine C-C motif ligand 8 (CCL8) in Escherichia coli (E. coli) has been achieved with approx. 1.5 mg protein/l culture. Quartz crystal microbalance (QCM) analysis exhibited that the purified CCL8 could bind with C-C chemokine receptor type 3 (CCR3) with dissociation equilibrium constant (K D) as 1.2 × 10-7 M in vitro. Obvious internalization of CCR3 in vivo could be detected in 1 h when exposed to 100 nM of CCL8. Compared with chemokine C-C motif ligand 11 (CCL11) and chemokine C-C motif ligand 24 (CCL24), a weaker chemotactic effect of CCR3 expressing cells was observed when induced by CCL8 with same concentration. CONCLUSION: This study delivers a simple and applicable way to produce functional chemokines in E. coli. The results clearly confirms that CCL8 can interact with chemokine receptor CCR3, therefore, it is promising area to develop drugs for the treatment of related diseases.
Assuntos
Quimiocina CCL8/metabolismo , Escherichia coli/genética , Receptores CCR3/agonistas , Quimiocina CCL8/genética , Quimiotaxia , Conjuntos de Dados como Assunto , Expressão Gênica , Células HEK293 , Humanos , Isopropiltiogalactosídeo , Ligantes , Plasmídeos , Ligação Proteica , Proteínas Recombinantes/genéticaRESUMO
Molecular fluorescence blinking provides a simple and attractive way to achieve super-resolution localization via conventional fluorescence microscopy. However, success in super-resolution imaging relies heavily on their blinking characteristics. We here report easily prepared and photostable nanoparticles, carbon dots (CDs), with desirable fluorescence blinking for high-density super-resolution imaging. The CDs exhibit a low duty cycle (â¼0.003) and high photon output (â¼8000) per switching event, as well as show much higher resistance to photobleaching than Alexa 647 or Cy5 typically used in single molecule localization microscopy. The stable blinking of CDs allows to perform high-density localization imaging at a resolution of 25 nm by sequentially recording the particle positions. The CD-based super-resolution imaging is further demonstrated by rendering CD-stained tubular peptide self-assemblies, CD-packed clusters with well-defined patterns, and CD-stained microtubules in a cell. Furthermore, this method has been validated as a valuable tool to detect the clustering and distribution of protein receptors in the plasma membrane that are not discerned with normal fluorescence imaging.
Assuntos
Carbono/química , Nanopartículas/química , Imagem Óptica/métodos , Membrana Celular/metabolismo , Células HeLa , Humanos , Espaço Intracelular/metabolismo , Fotodegradação , Razão Sinal-RuídoRESUMO
Revealing chemokine receptor CXCR4 expression, distribution, and internalization levels in different cancers helps to evaluate cancer progression or prognosis and to set personalized treatment strategy. We here describe a sensitive and high-throughput immunoassay for determining CXCR4 expression and distribution in cancer cells. The assay is accessible to a wide range of users in an ordinary lab only by dip-coating poly(styrene-co-N-isopropylacrylamide) spheres on the glass substrate. The self- assembled spheres form three-dimensional photonic colloidal crystals which enhance the fluorescence of CF647 and Alexa Fluor 647 by a factor of up to 1000. CXCR4 in cells is detected by using the sandwich immunoassay, where the primary antibody recognizes CXCR4 and the secondary antibody is labeled with CF647. With the newly established assay, we quantified the total expression of CXCR4, its distribution on the cell membrane and cytoplasm, and revealed their internalization level upon SDF-1α activation in various cancer cells, even for those with extremely low expression level.
Assuntos
Acrilamidas/química , Membrana Celular/metabolismo , Quimiocina CXCL12/química , Citoplasma/química , Receptores CXCR4/química , Estirenos/química , Linhagem Celular , Quimiocina CXCL12/metabolismo , Fluorescência , Humanos , Transdução de SinaisRESUMO
After the synthesis of transmembrane peptides/proteins (TMPs), their insertion into a lipid bilayer is a fundamental biophysical process. Moreover, correct orientations of TMPs in membranes determine the normal functions they play in relevant cellular activities. In this study, we have established a method to determine the orientation of TMPs in membranes. This method is based on the use of TAMRA, a fluorescent molecule with high extinction coefficient and fluorescence quantum yield, to act as a fluorescent probe and tryptophan as a quencher. Fluorescence quenching indicates that the model peptide displays membrane orientation with the N terminus outside and the C terminus inside dominantly. To elucidate the underlying mechanism, we performed molecular dynamics simulations. Our simulations suggest that both membrane insertion and the orientation of TMPs are determined by complex competition and cooperation between hydrophobic and electrostatic interactions. After initial membrane anchorage via electrostatic interactions of the charged residues with the lipid headgroups, further insertion is hindered by unfavorable interactions between the polar residues and lipid tails, which result in an energy barrier. Nevertheless, such a finite energy barrier is reduced by hydrophobic interactions between the non-polar residues and lipid tails. Moreover, a transient terminal flipping was captured to facilitate the membrane insertion. Once the inserted terminus reaches the opposite lipid headgroups, the hydrophobic interactions cooperate with the electrostatic interactions to complete the membrane insertion process.
Assuntos
Bicamadas Lipídicas/química , Simulação de Dinâmica Molecular , Peptídeos/química , Interações Hidrofóbicas e Hidrofílicas , Proteínas de Membrana/química , Espectrometria de Fluorescência , Triptofano/químicaRESUMO
Surfactants play a significant role in solubilization of photosystem I (PSI) in vitro. Triton X-100 (TX), n-Dodecyl-ß-D-maltoside (DDM), and sodium dodecyl sulfate (SDS) were employed to solubilize PSI particles in MES buffer to compare the effect of surfactant and its dosage on the apparent oxygen consumption rate of PSI. Through a combined assessment of sucrose density gradient centrifugation, Native PAGE and 77 K fluorescence with the apparent oxygen consumption, the nature of the enhancement of the apparent oxygen consumption activity of PSI by surfactants has been analyzed. Aggregated PSI particles can be dispersed by surfactant molecules into micelles, and the apparent oxygen consumption rate is higher for surfactant-solubilized PSI than for integral PSI particles. For DDM, PSI particles are solubilized mostly as the integral trimeric form. For TX, PSI particles are solubilized as incomplete trimeric and some monomeric forms. For the much harsher surfactant, SDS, PSI particles are completely solubilized as monomeric and its subunit forms. The enhancement of the oxygen consumption rate cannot be explained only by the effects of surfactant on the equilibrium between monomeric and trimeric forms of solubililized PSI. Care must be taken when the electron transfer activity of PSI is evaluated by methods based on oxygen consumption because the apparent oxygen consumption rate is influenced by uncoupled chlorophyll (Chl) from PSI, i.e., the larger the amount of uncoupled Chl, the higher the rate of apparent oxygen consumption. 77 K fluorescence spectra can be used to ensure that there is no uncoupled Chl present in the system. In order to eliminate the effect of trace uncoupled Chl, an efficient physical quencher of (1)O2, such as 1 mM NaN3, may be added into the mixture.
Assuntos
Consumo de Oxigênio/efeitos dos fármacos , Oxigênio/química , Complexo de Proteína do Fotossistema I/química , Complexo de Proteína do Fotossistema I/metabolismo , Spirulina/metabolismo , Tensoativos/química , Proteínas de Bactérias/química , Proteínas de Bactérias/metabolismo , Clorofila , Transferência de Energia , Consumo de Oxigênio/fisiologia , SolubilidadeRESUMO
A combination of abâ initio calculations, circular dichroism, nuclear magnetic resonance, and X-ray photoelectron spectroscopy has shown that aluminum ions can induce the formation of backbone ring structures in a wide range of peptides, including neurodegenerative disease related motifs. These ring structures greatly destabilize the protein and result in irreversible denaturation. This behavior benefits from the ability of aluminum ions to form chemical bonds simultaneously with the amide nitrogen and carbonyl oxygen atoms on the peptide backbone.
Assuntos
Alumínio/química , Proteínas/química , Teoria Quântica , Dicroísmo Circular , Espectroscopia de Ressonância Magnética , Conformação Proteica , Desnaturação ProteicaRESUMO
Algae, which are ubiquitous in ecosystems, have evolved a variety of light-harvesting complexes to better adapt to diverse habitats. Phycobilisomes/phycobiliproteins, unique to cyanobacteria, red algae, and certain cryptomonads, compensate for the lack of chlorophyll absorption, allowing algae to capture and efficiently transfer light energy in aquatic environments. With the advancement of microscopy and spectroscopy, the structure and energy transfer processes of increasingly complex phycobilisomes have been elucidated, providing us with a vivid portrait of the dynamic adaptation of their structures to the light environment in which algae thrive: 1) Cyanobacteria living on the surface of the water use short, small phycobilisomes to absorb red-orange light and reduce the damage from blue-violet light via multiple methods; 2) Large red algae inhabiting the depths of the ocean have evolved long and dense phycobilisomes containing phycoerythrin to capture the feeble blue-green light; 3) In far-red light environments such as caves, algae use special allophycocyanin cores to optimally utilize the far-red light; 4) When the environment shifts, algae can adjust the length, composition and density of their rods to better adapt; 5) By carefully designing the position of the pigments, phycobilisomes can transfer light energy to the reaction center with nearly 100% efficiency via three energy transfer processes.
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Cianobactérias , Ficobilissomas , Ficobilissomas/química , EcossistemaRESUMO
Spirulina platensis can secrete extracellular polymeric substances (EPS) helping to protect damage from stress environment, such as cadmium (Cd2+) exposure. However, the responding mechanism of S. platensis and the secreted EPS to exposure of Cd2+ is still unclear. This research focuses on the effects of Cd2+ on the composition and structure of the EPS and the response mechanism of EPS secretion from S. platensis for Cd2+ exposure. S. platensis can produce 261.37 mg·g-1 EPS when exposing to 20 mg·L-1 CdCl2, which was 2.5 times higher than the control group. The S. platensis EPS with and without Cd2+ treatment presented similar and stable irregularly fibrous structure. The monosaccharides composition of EPS in Cd2+ treated group are similar with control group but with different monosaccharides molar ratios, especially for Rha, Gal, Glc and Glc-UA. And the Cd2+ treatment resulted in a remarkable decline of humic acid and fulvic acid content. The antioxidant ability of S. platensis EPS increased significantly when exposed to 20 mg·L-1 CdCl2, which could be helpful for S. platensis protecting damage from high concentration of Cd2+. The transcriptome analysis showed that sulfur related metabolic pathways were up-regulated significantly, which promoted the synthesis of sulfur-containing amino acids and the secretion of large amounts of EPS.