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1.
Curr Issues Mol Biol ; 46(4): 3364-3378, 2024 Apr 14.
Artigo em Inglês | MEDLINE | ID: mdl-38666941

RESUMO

Neuroglobin (Ngb) is a cytosolic heme protein that plays an important role in protecting cells from apoptosis through interaction with oxidized cytochrome c (Cyt c) released from mitochondria. The interaction of reduced Ngb and oxidized Cyt c is accompanied by electron transfer between them and the reduction in Cyt c. Despite the growing number of studies on Ngb, the mechanism of interaction between Ngb and Cyt c is still unclear. Using Raman spectroscopy, we studied the effect of charged amino acid substitutions in Ngb and Cyt c on the conformation of their hemes. It has been shown that Ngb mutants E60K, K67E, K95E and E60K/E87K demonstrate changed heme conformations with the lower probability of the heme planar conformation compared to wild-type Ngb. Moreover, oxidized Cyt c mutants K25E, K72E and K25E/K72E demonstrate the decrease in the probability of methyl-radicals vibrations, indicating the higher rigidity of the protein microenvironment. It is possible that these changes can affect electron transfer between Ngb and Cyt c.

2.
Sci Adv ; 10(12): eadi2042, 2024 Mar 22.
Artigo em Inglês | MEDLINE | ID: mdl-38507479

RESUMO

The integration of heterogeneous modular units for building large-scale quantum networks requires engineering mechanisms that allow suitable transduction of quantum information. Magnon-based transducers are especially attractive due to their wide range of interactions and rich nonlinear dynamics, but most of the work to date has focused on linear magnon transduction in the traditional system composed of yttrium iron garnet and diamond, two materials with difficult integrability into wafer-scale quantum circuits. In this work, we present a different approach by using wafer-compatible materials to engineer a hybrid transducer that exploits magnon nonlinearities in a magnetic microdisc to address quantum spin defects in silicon carbide. The resulting interaction scheme points to the unique transduction behavior that can be obtained when complementing quantum systems with nonlinear magnonics.

3.
Nanomaterials (Basel) ; 14(6)2024 Mar 09.
Artigo em Inglês | MEDLINE | ID: mdl-38535643

RESUMO

Addressing periprosthetic infections, which present significant healing challenges that often require revision surgeries, necessitates the development of novel antibacterial materials and implants. Current research focuses on creating materials that hinder bacterial adhesion, colonization, and proliferation in surrounding tissues. Boron (B)-containing compounds are known for their antibacterial properties and potential in bone metabolism for regenerative medicine. In this study, we synthesized B-containing tricalcium phosphate (0.3B-TCP) with 1.1 wt.% B content via precipitation from aqueous solutions and sintering at 1100 °C. X-ray diffraction confirmed the ceramic's primary crystalline phase as ß-TCP, with B evenly distributed according to energy-dispersive spectroscopy data. Electron paramagnetic resonance (EPR) data verified stable paramagnetic borate anions, indicating successful BO33- substitution for phosphate groups. The microstructural properties of 0.3B-TCP ceramic were assessed before and after soaking in a saline solution. Its bending strength was approximately 30 MPa, and its porosity was about 33%. 0.3B-TCP ceramic demonstrated significant antimicrobial efficacy against various bacterial strains and a fungus. Cytotoxicity evaluation using equine adipose tissue-derived mesenchymal stem cells and osteogenic differentiation assessment were conducted. The combination of antibacterial efficacy and good cytocompatibility suggests 0.3B-TCP ceramic as a promising bone substitute material.

4.
J Fluoresc ; 2024 Mar 09.
Artigo em Inglês | MEDLINE | ID: mdl-38460095

RESUMO

The work presents a spectral-luminescent study of the sulfaguanidine transformation in water under a pulsed e-beam and UV irradiation of an UVb-04 bactericidal mercury lamp (from 180 to 275 nm), KrCl (222 nm), XeBr (282 nm) and XeCl (308 nm) excilamps. Fluorescent decay curves have been used in our analysis of the sulfaguanidine decomposition. The conversion of antibiotic under e-beam irradiation for up to 1 min was more than 80%, compared with UV radiation: UVb-04-26%, XeBr - 20%. KrCl and XeCl - about 10%. At the end of 64 min of irradiation with UVb-04 and XeBr lamps, the conversion was 99%. During irradiation with these lamps, sulfaguanidine almost completely decomposed and passed into the final fluorescent photoproducts. After e-beam irradiated at the end of 13 min the decrease in sulfaguanidine was 93%. At the same time, the formation of sulfaguanidine transformation products was minimal compared to UV irradiation. The effect of UV irradiation and a powerful e-beam on the decomposition mechanisms of sulfaguanidine are significantly different, which is manifested in various changes in the absorption and fluorescence spectra.

5.
J Photochem Photobiol B ; 252: 112870, 2024 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-38368635

RESUMO

Raman spectroscopy (RS), a powerful analytical technique, has gained increasing recognition and utility in the fields of biomedical and biological research. Raman spectroscopic analyses find extensive application in the field of medicine and are employed for intricate research endeavors and diagnostic purposes. Consequently, it enjoys broad utilization within the realm of biological research, facilitating the identification of cellular classifications, metabolite profiling within the cellular milieu, and the assessment of pigment constituents within microalgae. This article also explores the multifaceted role of RS in these domains, highlighting its distinct advantages, acknowledging its limitations, and proposing strategies for enhancement.


Assuntos
Análise Espectral Raman , Análise Espectral Raman/métodos
6.
Int J Biol Macromol ; 254(Pt 2): 127874, 2024 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-37939760

RESUMO

The Orange Carotenoid Protein (OCP) is a unique photoreceptor crucial for cyanobacterial photoprotection. Best studied Synechocystis sp. PCC 6803 OCP belongs to the large OCP1 family. Downregulated by the Fluorescence Recovery Protein (FRP) in low-light, high-light-activated OCP1 binds to the phycobilisomes and performs non-photochemical quenching. Recently discovered families OCP2 and OCP3 remain structurally and functionally underexplored, and no systematic comparative studies have ever been conducted. Here we present two first crystal structures of OCP2 from morphoecophysiologically different cyanobacteria and provide their comprehensive structural, spectroscopic and functional comparison with OCP1, the recently described OCP3 and all-OCP ancestor. Structures enable correlation of spectroscopic signatures with the effective number of hydrogen and discovered here chalcogen bonds anchoring the ketocarotenoid in OCP, as well as with the rotation of the echinenone's ß-ionone ring in the CTD. Structural data also helped rationalize the observed differences in OCP/FRP and OCP/phycobilisome functional interactions. These data are expected to foster OCP research and applications in optogenetics, targeted carotenoid delivery and cyanobacterial biomass engineering.


Assuntos
Proteínas de Bactérias , Synechocystis , Proteínas de Bactérias/química , Synechocystis/metabolismo , Análise Espectral , Carotenoides/química , Ficobilissomas/química
7.
Nanotechnology ; 35(15)2024 Jan 25.
Artigo em Inglês | MEDLINE | ID: mdl-38154127

RESUMO

Optically active point defects in semiconductors have received great attention in the field of solid-state quantum technologies. Hexagonal boron nitride, with an ultra-wide band gapEg= 6 eV, containing a negatively charged boron vacancy (VB-) with unique spin, optical, and coherent properties presents a new two-dimensional platform for the implementation of quantum technologies. This work establishes the value ofVB-spin polarization under optical pumping withλext= 532 nm laser using high-frequency (νmw= 94 GHz) electron paramagnetic resonance (EPR) spectroscopy. In optimal conditions polarization was found to beP≈ 38.4%. Our study reveals that Rabi oscillations induced on polarized spin states persist for up to 30-40µs, which is nearly two orders of magnitude longer than what was previously reported. Analysis of the coherent electron-nuclear interaction through the observed electron spin echo envelope modulation made it possible to detect signals from remote nitrogen and boron nuclei, and to establish a corresponding quadrupole coupling constantCq= 180 kHz related to nuclear quadrupole moment of14N. These results have fundamental importance for understanding the spin properties of boron vacancy.

8.
Biochim Biophys Acta Bioenerg ; 1865(1): 149014, 2024 01 01.
Artigo em Inglês | MEDLINE | ID: mdl-37739300

RESUMO

Phycobilisomes (PBSs) are giant water-soluble light-harvesting complexes of cyanobacteria and red algae, consisting of hundreds of phycobiliproteins precisely organized to deliver the energy of absorbed light to chlorophyll chromophores of the photosynthetic electron-transport chain. Quenching the excess of excitation energy is necessary for the photoprotection of photosynthetic apparatus. In cyanobacteria, quenching of PBS excitation is provided by the Orange Carotenoid Protein (OCP), which is activated under high light conditions. In this work, we describe parameters of anti-Stokes fluorescence of cyanobacterial PBSs in quenched and unquenched states. We compare the fluorescence readout from entire phycobilisomes and their fragments. The obtained results revealed the heterogeneity of conformations of chromophores in isolated phycobiliproteins, while such heterogeneity was not observed in the entire PBS. Under excitation by low-energy quanta, we did not detect a significant uphill energy transfer from the core to the peripheral rods of PBS, while the one from the terminal emitters to the bulk allophycocyanin chromophores is highly probable. We show that this direction of energy migration does not eliminate fluorescence quenching in the complex with OCP. Thus, long-wave excitation provides new insights into the pathways of energy conversion in the phycobilisome.


Assuntos
Cianobactérias , Ficobilissomas , Ficobilissomas/metabolismo , Proteínas de Bactérias/metabolismo , Fotossíntese , Cianobactérias/metabolismo , Espectrometria de Fluorescência/métodos
9.
Biochem Biophys Res Commun ; 683: 149119, 2023 11 26.
Artigo em Inglês | MEDLINE | ID: mdl-37862781

RESUMO

The Orange Carotenoid Protein (OCP) regulates cyanobacterial photosynthetic activity through photoactivation in intense light. A hydrogen bonding network involving the keto-carotenoid oxygen and Y201 and W288 residues prevents the spontaneous activation of dark-adapted OCP. To investigate the role of the hydrogen bonds in OCP photocycling, we introduced non-canonical amino acids near the keto-carotenoid, particularly iodine at the meta-position of Y201. This modification significantly increased the yield of red OCP photoproducts, albeit with a shorter lifetime. Changes in tryptophan fluorescence during photocycling influenced by the presence of iodine near W288 revealed interactions between Y201 and W288 in the absence of the carotenoid in the C-domain. We propose that upon the relaxation of red states, a ternary complex with the carotenoid is formed. Analysis of spectral signatures and interaction energies indicates that the specific iodo-tyrosine configuration enhances interactions between the carotenoid and W288.


Assuntos
Iodo , Triptofano , Aminoácidos , Ligação de Hidrogênio , Proteínas de Bactérias/metabolismo , Fluorescência , Luz , Carotenoides/metabolismo
10.
Inorg Chem ; 62(38): 15593-15604, 2023 Sep 25.
Artigo em Inglês | MEDLINE | ID: mdl-37695753

RESUMO

Technetium(I) 2 + 1 tricarbonyl complexes with a combination of N,N-bidentate ligands (2,2'-bipyridine, bipy; 1,10-phenanthroline, phen) and ethyl isocyanoacetate were prepared and characterized by NMR, IR, UV/visible, and luminescence spectroscopies and by high-performance liquid chromatography (HPLC). The crystal structures of [99Tc(CO)3(bipy)(CNCH2COOEt)](ClO4) (in the form of a solvate with 0.5CH2Cl2) and [99Tc(CO)3(phen)(CNCH2COOEt)](ClO4) (in the form of an adduct with an outer-sphere phen molecule) were determined by single-crystal X-ray diffraction. To evaluate the interfering effect of chloride ions on the formation of the 2 + 1 complexes, the kinetics of the replacement of labile monodentate ligand X in the complexes [MX(CO)3(N∧N)] (M = Re, 99Tc; N∧N = bipy, phen; X = Cl-, ClO4-) by CNCH2COOEt in ethanol were compared. The 99Tc bipy complexes with X = ClO4- (according to the IR data, perchlorate anion in ethanol is displaced from the coordination sphere by the solvent molecule) and X = Cl- are characterized by close ligand replacement rates. In the case of the 99Tc complexes with phen and Re complexes with both phen and bipy, the chloride complexes are appreciably less reactive than the chloride-free complexes. The technetium complexes are considerably more reactive in ligand replacement than their rhenium analogues. In the chloride-containing medium (saline), the complex [99mTc(CO)3(bipy)(CNCH2COOEt)]+ can be prepared under the conditions acceptable for nuclear medical applications, although higher isonitrile concentrations are required as compared to the chloride-free system.

11.
Int J Mol Sci ; 24(17)2023 Aug 31.
Artigo em Inglês | MEDLINE | ID: mdl-37686361

RESUMO

Hemoglobin is the main protein of red blood cells that provides oxygen transport to all cells of the human body. The ability of hemoglobin to bind the main low-molecular-weight thiol of the cell glutathione, both covalently and noncovalently, is not only an important part of the antioxidant protection of red blood cells, but also affects its affinity for oxygen in both cases. In this study, the properties of oxyhemoglobin in complex with reduced glutathione (GSH) and properties of glutathionylated hemoglobin bound to glutathione via an SS bond were characterized. For this purpose, the methods of circular dichroism, Raman spectroscopy, infrared spectroscopy, tryptophan fluorescence, differential scanning fluorimetry, and molecular modeling were used. It was found that the glutathionylation of oxyhemoglobin caused changes in the secondary structure of the protein, reducing the alpha helicity, but did not affect the heme environment, tryptophan fluorescence, and the thermostability of the protein. In the noncovalent complex of oxyhemoglobin with reduced glutathione, the secondary structure of hemoglobin remained almost unchanged; however, changes in the heme environment and the microenvironment of tryptophans, as well as a decrease in the protein's thermal stability, were observed. Thus, the formation of a noncovalent complex of hemoglobin with glutathione makes a more significant effect on the tertiary and quaternary structure of hemoglobin than glutathionylation, which mainly affects the secondary structure of the protein. The obtained data are important for understanding the functioning of glutathionylated hemoglobin, which is a marker of oxidative stress, and hemoglobin in complex with GSH, which appears to deposit GSH and release it during deoxygenation to increase the antioxidant protection of cells.


Assuntos
Antioxidantes , Oxiemoglobinas , Humanos , Triptofano , Hemoglobinas , Glutationa , Heme , Oxigênio
12.
PeerJ ; 11: e15781, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-37583913

RESUMO

From the Middle Jurassic to the Early Cretaceous, metriorhynchid crocodylomorphs inhabited marine ecosystems across the European archipelago. Unfortunately, European metriorhynchids are only well known from Germany, France, and the UK, with the Eastern European fossil record being especially poor. This hinders our understanding of metriorhynchid biodiversity across these continuous seaways, and our ability to investigate provincialism. Here we describe eleven isolated tooth crowns and six vertebrae referable to Metriorhynchidae from the Callovian, Oxfordian, Volgian (Tithonian), and Ryazanian (Berriasian) or Valanginian of European Russia. We also describe an indeterminate thalattosuchian tooth from the lower Bajocian of the Volgograd Oblast, the first discovery of a marine reptile from the Bajocian strata of European Russia. These rare fossils, along with previous reports of Russian thalattosuchians, indicate that thalattosuchians have been common in the Middle Russian Sea since it was formed. Palaeolatitude calculations for worldwide metriorhynchid-bearing localities demonstrate that the occurrences in European Russia are the most northern, located mainly between 44-50 degrees north. However, metriorhynchids appear to be rare at these palaeolatitudes, and are absent from palaeolatitudes higher than 50°. These observations support the hypothesis that metriorhynchids evolved an elevated metabolism but were not endo-homeothermic, especially as endo-homeothermic marine reptiles (ichthyosaurs and plesiosaurs) remained abundant at much higher palaeolatitudes.


Assuntos
Ecossistema , Dente , Animais , Fósseis , Répteis/anatomia & histologia , Biodiversidade
13.
Nat Commun ; 14(1): 5089, 2023 Aug 22.
Artigo em Inglês | MEDLINE | ID: mdl-37607945

RESUMO

Negatively-charged boron vacancy centers ([Formula: see text]) in hexagonal Boron Nitride (hBN) are attracting increasing interest since they represent optically-addressable qubits in a van der Waals material. In particular, these spin defects have shown promise as sensors for temperature, pressure, and static magnetic fields. However, their short spin coherence time limits their scope for quantum technology. Here, we apply dynamical decoupling techniques to suppress magnetic noise and extend the spin coherence time by two orders of magnitude, approaching the fundamental T1 relaxation limit. Based on this improvement, we demonstrate advanced spin control and a set of quantum sensing protocols to detect radiofrequency signals with sub-Hz resolution. The corresponding sensitivity is benchmarked against that of state-of-the-art NV-diamond quantum sensors. This work lays the foundation for nanoscale sensing using spin defects in an exfoliable material and opens a promising path to quantum sensors and quantum networks integrated into ultra-thin structures.

14.
Nature ; 619(7970): 539-544, 2023 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-37316662

RESUMO

Understanding the nature and origin of the precursor material to terrestrial planets is key to deciphering the mechanisms and timescales of planet formation1. Nucleosynthetic variability among rocky Solar System bodies can trace the composition of planetary building blocks2-5. Here we report the nucleosynthetic composition of silicon (µ30Si), the most abundant refractory planet-building element, in primitive and differentiated meteorites to identify terrestrial planet precursors. Inner Solar System differentiated bodies, including Mars, record µ30Si deficits of -11.0 ± 3.2 parts per million to -5.8 ± 3.0 parts per million whereas non-carbonaceous and carbonaceous chondrites show µ30Si excesses from 7.4 ± 4.3 parts per million to 32.8 ± 2.0 parts per million relative to Earth. This establishes that chondritic bodies are not planetary building blocks. Rather, material akin to early-formed differentiated asteroids must represent a major planetary constituent. The µ30Si values of asteroidal bodies correlate with their accretion ages, reflecting progressive admixing of a µ30Si-rich outer Solar System material to an initially µ30Si-poor inner disk. Mars' formation before chondrite parent bodies is necessary to avoid incorporation of µ30Si-rich material. In contrast, Earth's µ30Si composition necessitates admixing of 26 ± 9 per cent of µ30Si-rich outer Solar System material to its precursors. The µ30Si compositions of Mars and proto-Earth are consistent with their rapid formation by collisional growth and pebble accretion less than three million years after Solar System formation. Finally, Earth's nucleosynthetic composition for s-process sensitive (molybdenum and zirconium) and siderophile (nickel) tracers are consistent with pebble accretion when volatility-driven processes during accretion and the Moon-forming impact are carefully evaluated.

15.
Materials (Basel) ; 16(12)2023 Jun 20.
Artigo em Inglês | MEDLINE | ID: mdl-37374661

RESUMO

An alternative approach for the currently used replacement therapy in dentistry is to apply materials that restore tooth tissue. Among them, composites, based on biopolymers with calcium phosphates, and cells can be applied. In the present work, a composite based on polyvinylpyrrolidone (PVP) and alginate (Alg) with carbonate hydroxyapatite (CHA) was prepared and characterized. The composite was investigated by X-ray diffraction, infrared spectroscopy, electron paramagnetic resonance (EPR) and scanning electron microscopy methods, and the microstructure, porosity, and swelling properties of the material were described. In vitro studies included the MTT test using mouse fibroblasts, and adhesion and survivability tests with human dental pulp stem cells (DPSC). The mineral component of the composite corresponded to CHA with an admixture of amorphous calcium phosphate. The presence of a bond between the polymer matrix and CHA particles was shown by EPR. The structure of the material was represented by micro- (30-190 µm) and nano-pores (average 8.71 ± 4.15 nm). The swelling measurements attested that CHA addition increased the polymer matrix hydrophilicity by 200%. In vitro studies demonstrated the biocompatibility of PVP-Alg-CHA (95 ± 5% cell viability), and DPSC located inside the pores. It was concluded that the PVP-Alg-CHA porous composite is promising for dentistry applications.

16.
Materials (Basel) ; 16(7)2023 Apr 02.
Artigo em Inglês | MEDLINE | ID: mdl-37049124

RESUMO

Alginate is a natural polymer widely applied in materials science, medicine, and biotechnology. Its ability to bind metal ions in order to form insoluble gels has been comprehensively used to create capsules for cell technology, drug delivery, biomedical materials, etc. To modify and predict the properties of cross-linked alginate, knowledge about the mechanism of alginate binding with metal ions and the properties of its gels is necessary. This article presents the results obtained by proton Nuclear Magnetic Resonance Spectroscopy for alginate containing calcium and strontium (alkaline earth metal diamagnetic) ions and by Electron Paramagnetic Resonance Spectroscopy for alginate with copper (Cu) and manganese (Mn) (transition metal paramagnetic) ions. It was found that in the case of calcium (Ca) and Mn ions, their concentration does not affect their distribution in the alginate structure and the cross-linking density. In the case of strontium (Sr) and Cu ions, their number affects the number of binding sites and, accordingly, the cross-linking density. Thus, the cross-linking of alginate depends mainly on the characteristics of specific cations, while the nature of the bond (ionic or coordination type) is less important.

17.
Antioxidants (Basel) ; 12(2)2023 Feb 08.
Artigo em Inglês | MEDLINE | ID: mdl-36829973

RESUMO

Lipofuscin of retinal pigment epithelium (RPE) cells is a complex heterogeneous system of chromophores which accumulates as granules during the cell's lifespan. Lipofuscin serves as a source of various cytotoxic effects linked with oxidative stress. Several age-related eye diseases such as macular degeneration of the retina, as well as some severe inherited eye pathologies, are accompanied by a significant increase in lipofuscin granule concentration. The accumulation of carotenoids in the RPE could provide an effective antioxidant protection against lipofuscin cytotoxic manifestations. Given the highly lipophilic nature of carotenoids, their targeted delivery to the vulnerable tissues can potentially be assisted by special proteins. In this study, we demonstrate how protein-mediated delivery of zeaxanthin using water-soluble Bombyx mori carotenoid-binding protein (BmCBP-ZEA) suppresses the photoinducible oxidative stress in RPE cells caused by irradiation of lipofuscin with intense white light. We implemented fluorescence lifetime imaging of the RPE cell culture ARPE-19 fed with lipofuscin granules and then irradiated by white light with and without the addition of BmCBP-ZEA. We demonstrate that after irradiation the mean fluorescence lifetime of lipofuscin significantly increases, while the presence of BmCBP-ZEA at 200 nM concentration suppresses the increase in the average lifetime of lipofuscin fluorescence, indicating an approx. 35% inhibition of the oxidative stress. This phenomenon serves as indirect yet important evidence of the efficiency of the protein-mediated carotenoid delivery into pigment epithelium cells.

18.
ACS Nano ; 17(5): 4474-4482, 2023 Mar 14.
Artigo em Inglês | MEDLINE | ID: mdl-36802485

RESUMO

Semiconductor colloidal nanoplatelets based of CdSe have excellent optical properties. Their magneto-optical and spin-dependent properties can be greatly modified by implementing magnetic Mn2+ ions, using concepts well established for diluted magnetic semiconductors. A variety of magnetic resonance techniques based on high-frequency (94 GHz) electron paramagnetic resonance in continuous wave and pulsed mode were used to get detailed information on the spin structure and spin dynamics of Mn2+ ions in core/shell CdSe/(Cd,Mn)S nanoplatelets. We observed two sets of resonances assigned to the Mn2+ ions inside the shell and at the nanoplatelet surface. The surface Mn demonstrates a considerably longer spin dynamics than the inner Mn due to lower amount of surrounding Mn2+ ions. The interaction between surface Mn2+ ions and 1H nuclei belonging to oleic acid ligands is measured by means of electron nuclear double resonance. This allowed us to estimate the distances between the Mn2+ ions and 1H nuclei, which equal to 0.31 ± 0.04, 0.44 ± 0.09, and more than 0.53 nm. This study shows that the Mn2+ ions can serve as atomic-size probes for studying the ligand attachment to the nanoplatelet surface.

19.
J Phys Chem B ; 127(9): 1890-1900, 2023 03 09.
Artigo em Inglês | MEDLINE | ID: mdl-36799909

RESUMO

Most cyanobacteria utilize a water-soluble Orange Carotenoid Protein (OCP) to protect their light-harvesting complexes from photodamage. The Fluorescence Recovery Protein (FRP) is used to restore photosynthetic activity by inactivating OCP via dynamic OCP-FRP interactions, a multistage process that remains underexplored. In this work, applying time-resolved spectroscopy, we demonstrate that the interaction of FRP with the photoactivated OCP begins early in the photocycle. Interacting with the compact OCP state, FRP completely prevents the possibility of OCP domain separation and formation of the signaling state capable of interacting with the antenna. The structural element that prevents FRP binding and formation of the complex is the short α-helix at the beginning of the N-terminal domain of OCP, which masks the primary site in the C-terminal domain of OCP. We determined the rate of opening of this site and show that it remains exposed long after the relaxation of the red OCP states. Observations of the OCP transitions on the ms time scale revealed that the relaxation of the orange photocycle intermediates is accompanied by an increase in the interaction of the carotenoid keto group with the hydrogen bond donor tyrosine-201. Our data refine the current model of photoinduced OCP transitions and the interaction of its intermediates with FRP.


Assuntos
Proteínas de Bactérias , Cianobactérias , Proteínas de Bactérias/química , Cianobactérias/metabolismo , Análise Espectral , Transdução de Sinais , Carotenoides/química , Ficobilissomas/química
20.
Free Radic Biol Med ; 196: 133-144, 2023 02 20.
Artigo em Inglês | MEDLINE | ID: mdl-36649901

RESUMO

The balance between the mitochondrial respiratory chain activity and the cell's needs in ATP ensures optimal cellular function. Cytochrome c is an essential component of the electron transport chain (ETC), which regulates ETC activity, oxygen consumption, ATP synthesis and can initiate apoptosis. The impact of conformational changes in cytochrome c on its function is not understood for the lack of access to these changes in intact mitochondria. We have developed a novel sensor that uses unique properties of label-free surface-enhanced Raman spectroscopy (SERS) to identify conformational changes in heme of cytochrome c and to elucidate their role in functioning mitochondria. We have verified that molecule bond vibrations assessed by SERS are a reliable indicator of the heme conformation during changes in the inner mitochondrial membrane potential and ETC activity. We have demonstrated that cytochrome c heme reversibly switches between planar and ruffled conformations in response to the inner mitochondrial membrane potential (ΔΨ) and H+ concentration in the intermembrane space. This regulates the efficiency of the mitochondrial respiratory chain, thus, adjusting the mitochondrial respiration to the cell's consumption of ATP and the overall activity. We have found that under hypertensive conditions cytochrome c heme loses its sensitivity to ΔΨ that can affect the regulation of ETC activity. The ability of the proposed SERS-based sensor to track mitochondrial function opens broad perspectives in cell bioenergetics.


Assuntos
Citocromos c , Heme , Citocromos c/metabolismo , Heme/metabolismo , Potencial da Membrana Mitocondrial , Mitocôndrias/metabolismo , Trifosfato de Adenosina/metabolismo
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