Pub Date : 2022-11-15DOI: 10.29039/rusjbpc.2022.0511
A. Antonyan, A. Karapetyan, N. Petrosyan, P. Vardevanyan
Study on the interaction of DNA-specific compound Hoechst 33258 with bovine serum albumin (BSA) has been carried out, using the methods of absorption, differential and fluorescence spectroscopies. On the basis of the absorption and fluorescence spectra of the complexes of this ligand with BSA, in wide interval of the change of r (r=ligand/albumin), the dependence curves of the relative changes of the absorption (A/A0) and fluorescence (F/F0) maxima of the complexes H33258-BSA on the macromolecule concentration, were constructed. The dependence curve of A/A0 monotonously decreases along with protein concentration increasing due to the formation of the complexes. It was revealed that the dependence curve of F/F0 is not linear and is consisted of two regions: at low concentrations of the protein this curve decreases linearly, and passing through the minimum point, at relatively high concentrations of the protein starts linearly increasing. It was also shown that the differential spectra of the complexes H33258-BSA are characterized by hypo- and hyperchromic effects and hypsochromic shift: at low concentrations of the protein a hypochromic effect occurs, at higher concentrations of the protein (beginning from the certain ratios ligand/albumin ~1:1), the differential spectra are hypsochromically shifted with hyperchromic effect. The obtained data find out that Hoechst 33258, being DNA-specific ligand, tightly binds to albumin.
{"title":"SPECTRAL CHARACTERISTICS OF THE COMPLEXES OF HOECHST33258 WITH BSA","authors":"A. Antonyan, A. Karapetyan, N. Petrosyan, P. Vardevanyan","doi":"10.29039/rusjbpc.2022.0511","DOIUrl":"https://doi.org/10.29039/rusjbpc.2022.0511","url":null,"abstract":"Study on the interaction of DNA-specific compound Hoechst 33258 with bovine serum albumin (BSA) has been carried out, using the methods of absorption, differential and fluorescence spectroscopies. On the basis of the absorption and fluorescence spectra of the complexes of this ligand with BSA, in wide interval of the change of r (r=ligand/albumin), the dependence curves of the relative changes of the absorption (A/A0) and fluorescence (F/F0) maxima of the complexes H33258-BSA on the macromolecule concentration, were constructed. The dependence curve of A/A0 monotonously decreases along with protein concentration increasing due to the formation of the complexes. It was revealed that the dependence curve of F/F0 is not linear and is consisted of two regions: at low concentrations of the protein this curve decreases linearly, and passing through the minimum point, at relatively high concentrations of the protein starts linearly increasing. It was also shown that the differential spectra of the complexes H33258-BSA are characterized by hypo- and hyperchromic effects and hypsochromic shift: at low concentrations of the protein a hypochromic effect occurs, at higher concentrations of the protein (beginning from the certain ratios ligand/albumin ~1:1), the differential spectra are hypsochromically shifted with hyperchromic effect. The obtained data find out that Hoechst 33258, being DNA-specific ligand, tightly binds to albumin.","PeriodicalId":169374,"journal":{"name":"Russian Journal of Biological Physics and Chemisrty","volume":"93 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2022-11-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"115178306","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2022-11-15DOI: 10.29039/rusjbpc.2022.0513
A. Mammedzade, A. Mammadova, O. Gasymov
Nanoparticles are extensively used in various areas of industry. Among different nanoparticles, protein nanoparticles complexed with a wide range of drugs have a great potential for biomedical applications. Silk fibroin exhibits good biocompatibility properties and, therefore, is a good raw material for a wide variety of applications. In this study, structure and hydrophobic patch formation were studied in nanoparticles fabricated from silk fibroin. Far-UV circular dichroism spectroscopy and birefringence observed in a polarized microscope with Congo red staining indicate that fibroin nanoparticles are composed of small amyloid domains. Steady-state and time-resolved fluorescence of ANS revealed two hydrophobic patch formations. Decay-associated spectra of ANS bound to these patches show two species with lifetimes of about 4.2 ns and 14.8 ns. Dissociation constants for ANS complex formation for these patches are 8.3±0.4 M and 5.9±0.3 M, respectively. Acrylamide fluorescence quenching shows that solvent accessibility to native Trp residues is significantly decreased during fibroin nanoparticle formation. Data indicate that nanoparticles fabricated from fibroin are a good candidate for drug delivery applications.
{"title":"STRUCTURE OF SILK FIBROIN NANOPARTICLES: CHARACTERIZATION OF HYDROPHOBIC PATCHES","authors":"A. Mammedzade, A. Mammadova, O. Gasymov","doi":"10.29039/rusjbpc.2022.0513","DOIUrl":"https://doi.org/10.29039/rusjbpc.2022.0513","url":null,"abstract":"Nanoparticles are extensively used in various areas of industry. Among different nanoparticles, protein nanoparticles complexed with a wide range of drugs have a great potential for biomedical applications. Silk fibroin exhibits good biocompatibility properties and, therefore, is a good raw material for a wide variety of applications. In this study, structure and hydrophobic patch formation were studied in nanoparticles fabricated from silk fibroin. Far-UV circular dichroism spectroscopy and birefringence observed in a polarized microscope with Congo red staining indicate that fibroin nanoparticles are composed of small amyloid domains. Steady-state and time-resolved fluorescence of ANS revealed two hydrophobic patch formations. Decay-associated spectra of ANS bound to these patches show two species with lifetimes of about 4.2 ns and 14.8 ns. Dissociation constants for ANS complex formation for these patches are 8.3±0.4 M and 5.9±0.3 M, respectively. Acrylamide fluorescence quenching shows that solvent accessibility to native Trp residues is significantly decreased during fibroin nanoparticle formation. Data indicate that nanoparticles fabricated from fibroin are a good candidate for drug delivery applications.","PeriodicalId":169374,"journal":{"name":"Russian Journal of Biological Physics and Chemisrty","volume":"275 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2022-11-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"133103840","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2022-11-15DOI: 10.29039/rusjbpc.2022.0506
E. Sergeeva, M. Fadeeva, I. Khavronyuk, A. Mamontov, A. Ershov, A. Kuznetsov
Ctenophora are ancient and separate branch of evolutionary tree of the organic world development. They inhabit mostly epipelagic seas and demonstrate amazing mechanisms of adaptation to the environment. It is now possible with the potential technique of DNA sequencing to study biological objects not only experimentally but also theoretically. We studied opsins of the ctenophore Mnemiopsis leidyi A. Agassiz 1865 using bioinformatics methods. Additional annotation of opsins 1, 2, and 3, AFK83788.1, AFK83789.1, and AFK83790.1, respectively, was performed. Their 3D models were constructed and the localization of retinal within the protein molecules was determined. The protein environment of opsins was studied, and the first- and second-level protein neighbors were characterized. The structure of the protein-protein interaction network was shown to resemble a perceptron and, therefore, to perform the function of recognizing the patterns of the signals coming from outside. The multitasking of signaling pathways from endocytosis to Wnt signaling has been revealed, indicating the involvement of Ctenophora opsins in a wide range of genetic and physiological processes.
栉虫是有机世界发展进化树上古老而独立的分支。它们主要生活在上层海洋，并表现出惊人的适应环境的机制。随着DNA测序技术的发展，从理论上和实验上对生物对象进行研究已成为可能。采用生物信息学方法对栉水母leidyi A. Agassiz 1865的视蛋白进行了研究。分别对视蛋白1、2、3、AFK83788.1、AFK83789.1和AFK83790.1进行附加注释。他们建立了三维模型，并确定了视网膜在蛋白质分子中的定位。研究了视蛋白的蛋白质环境，并对其一级和二级邻接蛋白进行了表征。蛋白质-蛋白质相互作用网络的结构被证明类似于感知器，因此，执行识别来自外部信号模式的功能。从胞吞作用到Wnt信号通路的多任务信号通路已被揭示，表明视蛋白参与了广泛的遗传和生理过程。
{"title":"OPSINS OF THE CTENOPHORE MNEMIOPSIS LEIDYI AND A NETWORK OF PROTEIN-PROTEIN INTERACTIONS","authors":"E. Sergeeva, M. Fadeeva, I. Khavronyuk, A. Mamontov, A. Ershov, A. Kuznetsov","doi":"10.29039/rusjbpc.2022.0506","DOIUrl":"https://doi.org/10.29039/rusjbpc.2022.0506","url":null,"abstract":"Ctenophora are ancient and separate branch of evolutionary tree of the organic world development. They inhabit mostly epipelagic seas and demonstrate amazing mechanisms of adaptation to the environment. It is now possible with the potential technique of DNA sequencing to study biological objects not only experimentally but also theoretically. We studied opsins of the ctenophore Mnemiopsis leidyi A. Agassiz 1865 using bioinformatics methods. Additional annotation of opsins 1, 2, and 3, AFK83788.1, AFK83789.1, and AFK83790.1, respectively, was performed. Their 3D models were constructed and the localization of retinal within the protein molecules was determined. The protein environment of opsins was studied, and the first- and second-level protein neighbors were characterized. The structure of the protein-protein interaction network was shown to resemble a perceptron and, therefore, to perform the function of recognizing the patterns of the signals coming from outside. The multitasking of signaling pathways from endocytosis to Wnt signaling has been revealed, indicating the involvement of Ctenophora opsins in a wide range of genetic and physiological processes.","PeriodicalId":169374,"journal":{"name":"Russian Journal of Biological Physics and Chemisrty","volume":"1 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2022-11-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"130021231","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2022-11-15DOI: 10.29039/rusjbpc.2022.0518
O. Slatinskaya, G. Maksimov
The intracellular space of red blood cells (RBC) includes 40% hemoglobin (Hb) molecules and 60% being water molecules. When the intracellular volume changes, which is possible when erythrocyte cells pass through the blood stream, the possibility of redistribution of intracellular components of the cell due to its volume changes is assumed. By methods of optical spectroscopy (non-invasive methods of infrared spectroscopy, Raman-spectroscopy, laser interference microscopy) changes of morphology, conformation and redistribution of Hb have been revealed in the human erythrocyte due to an increase in the ratio of [Na+]in and [K+]in when Na+/K+-ATPase is blocked in the cell. The decrease of activity of Na+/K+-ATPase by a ouabaine (3 mM) has been found to lead not only to an increase in [Na+]in cell but also to an increase in positive charge on the cytoplasmic surface of the RBC membrane. In these conditions, changes in the conformation of both the heme and globine parts of the cytoplasmic Hb have been identified. It has been shown that cell depolarization, together with cell volume change, leads to a decrease in packing density of Hb molecule, which may be related to sorption of intracellular Na+ (or Ca2+) c Hb, as well as increasing the amount of water molecules in the cell and redistributin Hb in the cell. These processes can lead to a change in the conformation of Hb, as well as to a redistribution and alteration of the conformation of the cytoplasmic Hb.
红细胞(RBC)的胞内空间包括40%的血红蛋白(Hb)分子和60%的水分子。当细胞内体积发生变化时(红细胞通过血流时可能发生这种变化)，假设细胞内成分由于体积变化而重新分配的可能性。通过光学光谱方法(红外光谱、拉曼光谱、激光干涉显微镜等非侵入性方法)揭示了当Na+/K+- atp酶在细胞内被阻断时，由于[Na+]in和[K+]in的比例增加，红细胞中Hb的形态、构象和再分布发生了变化。研究发现，3 mM的瓦巴因降低Na+/K+- atp酶的活性，不仅导致细胞内[Na+]的增加，而且导致红细胞膜细胞质表面正电荷的增加。在这些条件下，已经确定了细胞质Hb的血红素和球蛋白部分构象的变化。研究表明，细胞去极化与细胞体积变化一起，导致Hb分子的堆积密度降低，这可能与细胞内Na+(或Ca2+) c Hb的吸附，以及细胞内水分子数量的增加和Hb在细胞内的重新分布有关。这些过程可导致血红蛋白构象的改变，以及细胞质血红蛋白构象的重新分布和改变。
{"title":"IN THE ERYTHROCYTE HEMOGLOBIN CONFORMATION AND DISTRIBUTION DURING RBC VOLUME CHANGES","authors":"O. Slatinskaya, G. Maksimov","doi":"10.29039/rusjbpc.2022.0518","DOIUrl":"https://doi.org/10.29039/rusjbpc.2022.0518","url":null,"abstract":"The intracellular space of red blood cells (RBC) includes 40% hemoglobin (Hb) molecules and 60% being water molecules. When the intracellular volume changes, which is possible when erythrocyte cells pass through the blood stream, the possibility of redistribution of intracellular components of the cell due to its volume changes is assumed. By methods of optical spectroscopy (non-invasive methods of infrared spectroscopy, Raman-spectroscopy, laser interference microscopy) changes of morphology, conformation and redistribution of Hb have been revealed in the human erythrocyte due to an increase in the ratio of [Na+]in and [K+]in when Na+/K+-ATPase is blocked in the cell. The decrease of activity of Na+/K+-ATPase by a ouabaine (3 mM) has been found to lead not only to an increase in [Na+]in cell but also to an increase in positive charge on the cytoplasmic surface of the RBC membrane. In these conditions, changes in the conformation of both the heme and globine parts of the cytoplasmic Hb have been identified. It has been shown that cell depolarization, together with cell volume change, leads to a decrease in packing density of Hb molecule, which may be related to sorption of intracellular Na+ (or Ca2+) c Hb, as well as increasing the amount of water molecules in the cell and redistributin Hb in the cell. These processes can lead to a change in the conformation of Hb, as well as to a redistribution and alteration of the conformation of the cytoplasmic Hb.","PeriodicalId":169374,"journal":{"name":"Russian Journal of Biological Physics and Chemisrty","volume":"40 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2022-11-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"116052372","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2022-11-15DOI: 10.29039/rusjbpc.2022.0514
T. Rybinskaya, O. Portnyagina, E. Zelepuga, V. Khomenko, N. Kim, E. Chingizova, E. Menchinskaya, V. Glazunov, D. Chistyulin, O. Novikova
The paper considers the process of formation and properties of amyloid-like aggregates of outer membrane non-specific porins (OmpC and OmpF) of the gram-negative bacterium Yersinia pseudotuberculos in an acidic medium (pH 4.5) at elevated temperature. The dynamics of the formation of amyloid-like aggregates of porins was monitored after two and four weeks of incubation (at 42 °C) and after 3-5 hours (at 90 °C) by staining the samples with amyloid-specific dye thioflavin T, analyzing the spectra of circular dichroism in the far UV region, IR -spectroscopy and confocal microscopy. It was found that in the case of porin OmpC, incubation under mild conditions (42°C) leads to a reversible accumulation of α-helical regions in the protein polypeptide chain. No significant changes are observed in the spatial structure of OmpF porin under these conditions, however, under harsh conditions (95 ºC) amyloid-like aggregates are formed, which are characterized by an increased content of the β-sheet structure. Using IR spectroscopy, it was shown that the conformational rearrangement in the molecule of OmpF porin is associated with a change in the quantity and quality of elements of the β-structure. According to confocal microscopy, the aggregates of the studied non-specific porins can be considered as intermediate products of the amyloidogenic pathway - oligomers. According to the literature data, these oligomers, which precede the formation of mature fibrils, have membranolytic and cytotoxic properties. For heated samples of the studied porins during reconstitution into bilayer lipid membranes, neither pore-forming nor membranolytic activity was detected. With respect to Neuro-2a CCL-131™ mouse neuroblastoma cells, the aggregates of OmpF and OmpC porins obtained after incubation had a higher toxicity compared to the initial protein samples.
{"title":"STUDY ON THE AMYLOIDOIGENIC POTENTIAL OF NON-SPECIFIC YERSINIA PSEUDOTUBERCULOSIS PORINS","authors":"T. Rybinskaya, O. Portnyagina, E. Zelepuga, V. Khomenko, N. Kim, E. Chingizova, E. Menchinskaya, V. Glazunov, D. Chistyulin, O. Novikova","doi":"10.29039/rusjbpc.2022.0514","DOIUrl":"https://doi.org/10.29039/rusjbpc.2022.0514","url":null,"abstract":"The paper considers the process of formation and properties of amyloid-like aggregates of outer membrane non-specific porins (OmpC and OmpF) of the gram-negative bacterium Yersinia pseudotuberculos in an acidic medium (pH 4.5) at elevated temperature. The dynamics of the formation of amyloid-like aggregates of porins was monitored after two and four weeks of incubation (at 42 °C) and after 3-5 hours (at 90 °C) by staining the samples with amyloid-specific dye thioflavin T, analyzing the spectra of circular dichroism in the far UV region, IR -spectroscopy and confocal microscopy. It was found that in the case of porin OmpC, incubation under mild conditions (42°C) leads to a reversible accumulation of α-helical regions in the protein polypeptide chain. No significant changes are observed in the spatial structure of OmpF porin under these conditions, however, under harsh conditions (95 ºC) amyloid-like aggregates are formed, which are characterized by an increased content of the β-sheet structure. Using IR spectroscopy, it was shown that the conformational rearrangement in the molecule of OmpF porin is associated with a change in the quantity and quality of elements of the β-structure. According to confocal microscopy, the aggregates of the studied non-specific porins can be considered as intermediate products of the amyloidogenic pathway - oligomers. According to the literature data, these oligomers, which precede the formation of mature fibrils, have membranolytic and cytotoxic properties. For heated samples of the studied porins during reconstitution into bilayer lipid membranes, neither pore-forming nor membranolytic activity was detected. With respect to Neuro-2a CCL-131™ mouse neuroblastoma cells, the aggregates of OmpF and OmpC porins obtained after incubation had a higher toxicity compared to the initial protein samples.","PeriodicalId":169374,"journal":{"name":"Russian Journal of Biological Physics and Chemisrty","volume":"4 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2022-11-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"123706088","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2022-11-15DOI: 10.29039/rusjbpc.2022.0508
K. Tereshkina, N. Loiko, E. Tereshkin, V. Kovalenko, A. Generalova, Y. Krupyanskii
The formation of complexes and crystals of bacterial nucleoid DNA inside bacterial cells in response to adverse external influences is of great interest both in biophysics and structural biology, and in various fields of industry. The main role in the condensation and crystallization of DNA in bacteria under stressful conditions (starvation, temperature, oxidative, and other types of stress) is played by DNA-binding proteins DPS. In this work, the dynamic behavior and structure of Escherichia coli DPS protein complexes in solution with short-chain DNA (25 base pairs) are studied using classical molecular dynamics in the all-atom approximation with a change in the concentration of the phenolic lipid 4-hexylresorcinol, which is a chemical analog of bacterial anabiosis inducers. The effect of changing the concentration of 4-hexylresorcinol from 0 to 50, 100 and 500 molecules on the DNA-DPS complex is being studied. By searching for the linear interaction energy, data were obtained on the free energies of DNA-protein binding without and in the presence of 4-hexylresorcinol. It has been shown that high concentrations of 4-hexylresorcinol promote the formation of DNA complexes with DPS. To identify the features of the dynamic behavior of DNA and protein, the principal component analysis was carried out. The spatial and energy characteristics of the complexes was obtained.
{"title":"STRUCTURAL PECULIARITIES OF DNA-DPS CLUSTERS AT DIFFERENT CONCENTRATIONS OF 4-HEXYLRESORCINOL","authors":"K. Tereshkina, N. Loiko, E. Tereshkin, V. Kovalenko, A. Generalova, Y. Krupyanskii","doi":"10.29039/rusjbpc.2022.0508","DOIUrl":"https://doi.org/10.29039/rusjbpc.2022.0508","url":null,"abstract":"The formation of complexes and crystals of bacterial nucleoid DNA inside bacterial cells in response to adverse external influences is of great interest both in biophysics and structural biology, and in various fields of industry. The main role in the condensation and crystallization of DNA in bacteria under stressful conditions (starvation, temperature, oxidative, and other types of stress) is played by DNA-binding proteins DPS. In this work, the dynamic behavior and structure of Escherichia coli DPS protein complexes in solution with short-chain DNA (25 base pairs) are studied using classical molecular dynamics in the all-atom approximation with a change in the concentration of the phenolic lipid 4-hexylresorcinol, which is a chemical analog of bacterial anabiosis inducers. The effect of changing the concentration of \u00004-hexylresorcinol from 0 to 50, 100 and 500 molecules on the DNA-DPS complex is being studied. By searching for the linear interaction energy, data were obtained on the free energies of DNA-protein binding without and in the presence of 4-hexylresorcinol. It has been shown that high concentrations of \u00004-hexylresorcinol promote the formation of DNA complexes with DPS. To identify the features of the dynamic behavior of DNA and protein, the principal component analysis was carried out. The spatial and energy characteristics of the complexes was obtained.","PeriodicalId":169374,"journal":{"name":"Russian Journal of Biological Physics and Chemisrty","volume":"70 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2022-11-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"131109175","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2022-11-15DOI: 10.29039/rusjbpc.2022.0522
N. Ronzhin, E. Posokhina, O. Mogilnaya, A. Puzyr, J. Gitelson, V. Bondar
The paper presents data that testify in favor of the participation of the cytochrome P450 system in the light emission of higher fungi. Extracts from mycelia of different species of luminous basidiomycetes containing fungal luminescent systems that provide luminescence in vitro were obtained. Applied conditions for the isolation of luminescent systems (sonication, centrifugation at 40000g) indicate the presence of membrane structures in the extracts, in particular, microsomes formed as a result of ultrasonic disintegration of the endoplasmic reticulum (ER). Differential spectral analysis of the extracts revealed the presence of two absorption peaks at 410 nm and 450 nm, which indicates the presence of cytochromes b5 and P450. The luminescence of the extracts is stimulated by reduced pyridine nucleotides, however, the addition of NADPH causes a higher level of luminescence compared with NADH. The addition of hydrogen peroxide significantly (from several times to 1-2 orders of magnitude) increases the luminescence intensity of extracts activated by NAD(P)H. The addition of fluconazole significantly inhibits the light emission of extracts. The data obtained indicates that the cytochrome P450 system associated with ER membranes may participate in the mechanism of light emission of higher fungi with the involvement in the process of electron transport enzyme systems: NADPH-dependent reductase of cytochrome P450 - cytochrome P450 and NADH-dependent reductase of cytochrome b5 - cytochrome b5 - cytochrome P450. In this case, cytochrome P450 may hydroxylate hispidin (precursor of the luminescent reaction substrate) to form luciferin and catalyze its oxidation in the presence of ROS with light emission.
{"title":"CYTOCHROME P450 SYSTEM MAY BE INVOLVED IN THE LIGHT EMISSION OF HIGHER FUNGI","authors":"N. Ronzhin, E. Posokhina, O. Mogilnaya, A. Puzyr, J. Gitelson, V. Bondar","doi":"10.29039/rusjbpc.2022.0522","DOIUrl":"https://doi.org/10.29039/rusjbpc.2022.0522","url":null,"abstract":"The paper presents data that testify in favor of the participation of the cytochrome P450 system in the light emission of higher fungi. Extracts from mycelia of different species of luminous basidiomycetes containing fungal luminescent systems that provide luminescence in vitro were obtained. Applied conditions for the isolation of luminescent systems (sonication, centrifugation at 40000g) indicate the presence of membrane structures in the extracts, in particular, microsomes formed as a result of ultrasonic disintegration of the endoplasmic reticulum (ER). Differential spectral analysis of the extracts revealed the presence of two absorption peaks at 410 nm and 450 nm, which indicates the presence of cytochromes b5 and P450. The luminescence of the extracts is stimulated by reduced pyridine nucleotides, however, the addition of NADPH causes a higher level of luminescence compared with NADH. The addition of hydrogen peroxide significantly (from several times to 1-2 orders of magnitude) increases the luminescence intensity of extracts activated by NAD(P)H. The addition of fluconazole significantly inhibits the light emission of extracts. The data obtained indicates that the cytochrome P450 system associated with ER membranes may participate in the mechanism of light emission of higher fungi with the involvement in the process of electron transport enzyme systems: NADPH-dependent reductase of cytochrome P450 - cytochrome P450 and NADH-dependent reductase of cytochrome b5 - cytochrome b5 - cytochrome P450. In this case, cytochrome P450 may hydroxylate hispidin (precursor of the luminescent reaction substrate) to form luciferin and catalyze its oxidation in the presence of ROS with light emission.","PeriodicalId":169374,"journal":{"name":"Russian Journal of Biological Physics and Chemisrty","volume":"49 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2022-11-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"129664995","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2022-11-15DOI: 10.29039/rusjbpc.2022.0516
A. Litvinenko, V. Nekrasov, E. Chetvertak, V. Maltsev
Cardiovascular diseases occupy a leading place among the causes of death in the population. The main complications of the disease in this group are closely related to changes in the functioning of the hemostasis system, including primary hemostasis. Antiplatelet therapy is used to prevent the development of complications associated with disorders in the hemostasis system. This paper examines the behavior of the platelet population of healthy donors before and after the use of two main types of drugs: acetylsalicylic acid and clopidogrel. Using the method of scanning flow cytometry, the shape index of single platelets was measured, followed by the construction of distributions according to this characteristic for native samples and after exposure to a small amount of an activation agonist. The sensitivity of platelets to the activation agonist is assessed by the change in distributions before and after the application of a short course of therapy. As a result of the assessment, the threshold value for the onset of changes in the shape of platelets and sensitivity increased with the use of acetylsalicylic acid, and decreased with the use of clopidogrel. The demonstrated approach for assessing sensitivity can be useful both in assessing the degree of effectiveness of antiplatelet therapy used and in diagnosing the presence of complications in the course of diseases of the cardiovascular system on the part of primary hemostasis.
{"title":"EVALUATION OF THE HUMAN BLOOD PLATELETS SENSITIVITY TO AN ACTIVATION AGONIST ADENOSINE DIPHOSPHATE BY SCANNING FLOW CYTOMETRY UNDER CONDITIONS OF ANTIPLATELET THERAPY","authors":"A. Litvinenko, V. Nekrasov, E. Chetvertak, V. Maltsev","doi":"10.29039/rusjbpc.2022.0516","DOIUrl":"https://doi.org/10.29039/rusjbpc.2022.0516","url":null,"abstract":"Cardiovascular diseases occupy a leading place among the causes of death in the population. The main complications of the disease in this group are closely related to changes in the functioning of the hemostasis system, including primary hemostasis. Antiplatelet therapy is used to prevent the development of complications associated with disorders in the hemostasis system. This paper examines the behavior of the platelet population of healthy donors before and after the use of two main types of drugs: acetylsalicylic acid and clopidogrel. Using the method of scanning flow cytometry, the shape index of single platelets was measured, followed by the construction of distributions according to this characteristic for native samples and after exposure to a small amount of an activation agonist. The sensitivity of platelets to the activation agonist is assessed by the change in distributions before and after the application of a short course of therapy. As a result of the assessment, the threshold value for the onset of changes in the shape of platelets and sensitivity increased with the use of acetylsalicylic acid, and decreased with the use of clopidogrel. The demonstrated approach for assessing sensitivity can be useful both in assessing the degree of effectiveness of antiplatelet therapy used and in diagnosing the presence of complications in the course of diseases of the cardiovascular system on the part of primary hemostasis.","PeriodicalId":169374,"journal":{"name":"Russian Journal of Biological Physics and Chemisrty","volume":"33 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2022-11-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"121152733","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2022-11-15DOI: 10.29039/rusjbpc.2022.0502
G. Agaeva, G. Safarli, N. Godjaev
The features of the spatial organization of the hylambatin molecule were investigated by methods of molecular mechanics and molecular dynamics. Hylambatin consists of twelve amino acid residues in the sequence: Asp-Pro-Pro-Asp-Pro-Asn-Arg-Phe-Tyr-Gly-Met-Met-NH2. Unlike all other tachykinins, hylambatin has a Met residue replacing the usual Leu at penultimate position. The tachykinin peptide hylambatin has been isolated and chemically characterized from methanol extracts of the skin of Hylambates maculatus, an African rhacophorid frog. It has been shown that intravenously administered hylambatin significantly increases the level of glucose and insulin in blood plasma. In this paper, the conformational flexibility of the hylambatin molecule was studied by methods of molecular mechanics and molecular dynamics. The conformational calculation of the peptide took into account non-valent and electrostatic interactions, hydrogen bonds and torsion potentials. Based on fragmentary analysis, stable spatial structures of the hylambatin dodecapeptide were determined, which can be represented as a set of conformations characterized by a relatively labile N-terminal tetrapeptide and a conformationally rigid C-terminal octapeptide. In the calculated stable conformational states, the effective interactions of the side chains of residues and hydrogen bonds were refined and energetically evaluated. It has been shown that the hylambatin molecule preferably forms practically isoenergetic conformations with various structural types at the N-end of the peptide chain, passing into the alpha helix at the C-end. By the method of molecular dynamics, the pattern of intramolecular mobility of stable conformations of the hylambatin molecule was modeled both in vacuum and surrounded by water molecules. Based on the calculated values of the dihedral angles, molecular models of energetically preferred conformational states of the hylambatin dodecapeptide were constructed.
采用分子力学和分子动力学方法研究了海兰蛋白分子的空间组织特征。hylamatin由12个氨基酸残基组成，序列为:asp - pro - pro - asp - pro - asn - arg - ph - tyr - gly - met - met - nh2。与所有其他速激素不同，hylamatin在倒数第二的位置有一个Met残基取代通常的Leu。从非洲棘足蛙(hylamates maculatus)皮肤的甲醇提取物中分离出速激肽(hylamatin)，并对其进行了化学表征。研究表明，静脉注射hylamatin可显著提高血浆中葡萄糖和胰岛素的水平。本文采用分子力学和分子动力学的方法研究了hylamatin分子的构象柔韧性。肽的构象计算考虑了非价相互作用和静电相互作用、氢键和扭转势。基于片段分析，确定了hylamatin十二肽的稳定空间结构，可以表示为相对不稳定的n端四肽和构象刚性的c端八肽的一组构象。在计算的稳定构象态中，对残基侧链和氢键的有效相互作用进行了细化和能量评价。研究表明，hylambatin分子在肽链的n端形成具有各种结构类型的等能构象，并在c端进入α螺旋。利用分子动力学的方法，模拟了hylamatin分子在真空和水分子包围下稳定构象的分子内迁移模式。基于二面角的计算值，构建了hylamatin十二肽的能量优选构象的分子模型。
{"title":"MOLECULAR MODELLİNG OF CONFORMATİONAL FLEXİBİLİTY OF HYLAMBATİN MOLECULE","authors":"G. Agaeva, G. Safarli, N. Godjaev","doi":"10.29039/rusjbpc.2022.0502","DOIUrl":"https://doi.org/10.29039/rusjbpc.2022.0502","url":null,"abstract":"The features of the spatial organization of the hylambatin molecule were investigated by methods of molecular mechanics and molecular dynamics. Hylambatin consists of twelve amino acid residues in the sequence: Asp-Pro-Pro-Asp-Pro-Asn-Arg-Phe-Tyr-Gly-Met-Met-NH2. Unlike all other tachykinins, hylambatin has a Met residue replacing the usual Leu at penultimate position. The tachykinin peptide hylambatin has been isolated and chemically characterized from methanol extracts of the skin of Hylambates maculatus, an African rhacophorid frog. It has been shown that intravenously administered hylambatin significantly increases the level of glucose and insulin in blood plasma. In this paper, the conformational flexibility of the hylambatin molecule was studied by methods of molecular mechanics and molecular dynamics. The conformational calculation of the peptide took into account non-valent and electrostatic interactions, hydrogen bonds and torsion potentials. Based on fragmentary analysis, stable spatial structures of the hylambatin dodecapeptide were determined, which can be represented as a set of conformations characterized by a relatively labile N-terminal tetrapeptide and a conformationally rigid C-terminal octapeptide. In the calculated stable conformational states, the effective interactions of the side chains of residues and hydrogen bonds were refined and energetically evaluated. It has been shown that the hylambatin molecule preferably forms practically isoenergetic conformations with various structural types at the N-end of the peptide chain, passing into the alpha helix at the C-end. By the method of molecular dynamics, the pattern of intramolecular mobility of stable conformations of the hylambatin molecule was modeled both in vacuum and surrounded by water molecules. Based on the calculated values of the dihedral angles, molecular models of energetically preferred conformational states of the hylambatin dodecapeptide were constructed.","PeriodicalId":169374,"journal":{"name":"Russian Journal of Biological Physics and Chemisrty","volume":"1 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2022-11-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"131304274","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2022-11-15DOI: 10.29039/rusjbpc.2022.0520
E. Timokhina, N. Yaglova, S. Obernikhin, V. Yaglov
The ability to reduce the isotopic composition of hydrogen in the internal environment of the body to influence the physiology of the organs of the endocrine system is a new and practically unexplored direction in science. The aim of the work was to study the morphological and functional changes in the thyroid gland of male rats after a two-week decrease in the content of deuterium in the internal environment of the body. The experiment was carried out on male Wistar rats fed deuterium-reduced water instead of drinking water for 2 weeks. The main effect exerted by water with a reduced content of deuterium on the body is a gradual decrease in the content of deuterium in liquid media and tissues due to isotope exchange reactions. Two weeks after the start of the experiment, the concentrations of thyroid hormones were determined: total and free thyroxine, total and free triiodothyronine, and thyroid-stimulating hormone. Histological and morphometric examination of thyroid preparations was carried out. Morphological and functional changes in the thyroid gland of male rats after a two-week reduction of the deuterium content in the internal environment of the body was studied. A prolonged decrease in deuterium influx led to structural changes in the thyroid gland and its secretory activity and were associated with hypothalamic-pituitary hypothyroidism due to a decrease in the synthesis of thyroid-stimulating hormone.
{"title":"CHANGES IN THYROID MORPHOLOGY AND FUNCTION PRODUCED BY THE SHIFT IN DEUTERIUM/PROTIUM BALANCE IN THE ORGANISM","authors":"E. Timokhina, N. Yaglova, S. Obernikhin, V. Yaglov","doi":"10.29039/rusjbpc.2022.0520","DOIUrl":"https://doi.org/10.29039/rusjbpc.2022.0520","url":null,"abstract":"The ability to reduce the isotopic composition of hydrogen in the internal environment of the body to influence the physiology of the organs of the endocrine system is a new and practically unexplored direction in science. The aim of the work was to study the morphological and functional changes in the thyroid gland of male rats after a two-week decrease in the content of deuterium in the internal environment of the body. The experiment was carried out on male Wistar rats fed deuterium-reduced water instead of drinking water for 2 weeks. The main effect exerted by water with a reduced content of deuterium on the body is a gradual decrease in the content of deuterium in liquid media and tissues due to isotope exchange reactions. Two weeks after the start of the experiment, the concentrations of thyroid hormones were determined: total and free thyroxine, total and free triiodothyronine, and thyroid-stimulating hormone. Histological and morphometric examination of thyroid preparations was carried out. Morphological and functional changes in the thyroid gland of male rats after a two-week reduction of the deuterium content in the internal environment of the body was studied. A prolonged decrease in deuterium influx led to structural changes in the thyroid gland and its secretory activity and were associated with hypothalamic-pituitary hypothyroidism due to a decrease in the synthesis of thyroid-stimulating hormone.","PeriodicalId":169374,"journal":{"name":"Russian Journal of Biological Physics and Chemisrty","volume":"95 20 Supplement 1 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2022-11-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"128056448","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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