Biochemistry (Moscow), Supplement Series A: Membrane and Cell Biology最新文献

Comparison of Dopamine and Tyramine Action on the Firing Rate of Trigeminal Dural Afferents in Wild Type and DAT-HET Rats 多巴胺和酪胺对野生型和DAT-HET大鼠三叉神经硬脑膜传入事件放电率的影响

IF 1.1

Biochemistry (Moscow), Supplement Series A: Membrane and Cell Biology Pub Date : 2025-04-24 DOI: 10.1134/S1990747824700533

S. Svitko, K. Shaidullova, E. Nevsky, A. Ananev, G. Sitdikova

{"title":"Comparison of Dopamine and Tyramine Action on the Firing Rate of Trigeminal Dural Afferents in Wild Type and DAT-HET Rats","authors":"S. Svitko, K. Shaidullova, E. Nevsky, A. Ananev, G. Sitdikova","doi":"10.1134/S1990747824700533","DOIUrl":"10.1134/S1990747824700533","url":null,"abstract":"Migraine is a widespread disorder which is defined by persistent and recurring headache attacks. Trigeminal-vascular system (TVS), including meningeal afferents of trigeminal ganglion, is considered to be a key structure in generating nociceptive signals in migraine, in which dopaminergic and trace amine-mediated effects remain unexplored. DAT-HET rats, heterozygous of DAT-KO line, provide a useful model for studying the role of moderate changes in DA homeostasis in migraine mechanisms. In this research, we investigated tyramine and dopamine (DA) effects in trigeminal peripheral afferents with elecrophysiological recording of action potentials of the trigeminal nerve in the rat hemiskull preparations. The data obtained indicates that DA application exerts pro-nociceptive action in trigeminal meningeal afferents in both WT (wild type) and DAT-HET groups in similar manner. Tyramine application also exhibited pro-nociceptive action in peripheral afferents, but demonstrated prolonged pro-nociceptive effect in DAT-HET group. Our data contributes to understanding of dopaminergic signaling in TVS by functionally confirming interplay between dopaminergic system and trace amines in peripheral afferents; as well as by demonstrating of novel electrophysiological evidence of tyramine pro-nociceptive action in TVS.","PeriodicalId":484,"journal":{"name":"Biochemistry (Moscow), Supplement Series A: Membrane and Cell Biology","volume":"19 1","pages":"106 - 114"},"PeriodicalIF":1.1,"publicationDate":"2025-04-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143865558","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}

引用次数: 0

Behavioral Correlates of Migraine, Oxidative Stress, and CGRP Levels in the Experimental Model of Chronic Migraine with Aura in Rats 大鼠先兆性慢性偏头痛实验模型中偏头痛的行为相关性、氧化应激和 CGRP 水平

IF 1.1

Biochemistry (Moscow), Supplement Series A: Membrane and Cell Biology Pub Date : 2025-04-24 DOI: 10.1134/S1990747824700417

K. Bogatova, A. Yakovlev, G. Sitdikova, O. Yakovleva

{"title":"Behavioral Correlates of Migraine, Oxidative Stress, and CGRP Levels in the Experimental Model of Chronic Migraine with Aura in Rats","authors":"K. Bogatova, A. Yakovlev, G. Sitdikova, O. Yakovleva","doi":"10.1134/S1990747824700417","DOIUrl":"10.1134/S1990747824700417","url":null,"abstract":"Migraine is a common chronic neurovascular disease accompanied by unilateral headache, photophobia, allodynia, and autonomic disturbances. About 1/3 of migraine patients have a transient neurological deficit that precedes or overlaps the headache phase, called aura. The main physiological mechanism underlying aura is thought to be cortical spreading depression (CSD), a wave of massive depolarization passing through neurons and glial cells of the cortex. The aim of the present study was to develop the model of chronic migraine with aura in rats and analyze the main behavioral and biochemical correlates of migraine. Male rats were subjected to surgery with the installation of a mini bath providing access to the dura mater above the somatosensory cortex. KCl was applied on the dura mater to induce CSD every other day for 9 days. Sham rats obtained NaCl application according to the same protocol. Behavioral tests were performed before and after application of KCl or NaCl. Repeated KCl application decreased locomotor and exploratory activity, as well as cognitive ability, increased anxiety and induced photophobia and allodynia in rats. Besides, rats with a model of chronic migraine with aura showed increased plasma calcitonin gene-related peptide (CGRP) concentration and decreased glutathione peroxidase activity in brain tissue. In conclusion, the proposed model can be used to study the behavioral correlates of chronic migraine with aura in experimental animals, which will allow us to investigate the underlying mechanisms and develop treatment approaches.","PeriodicalId":484,"journal":{"name":"Biochemistry (Moscow), Supplement Series A: Membrane and Cell Biology","volume":"19 1","pages":"27 - 34"},"PeriodicalIF":1.1,"publicationDate":"2025-04-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143865546","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}

引用次数: 0

Studying Ferritin Self-Assembly Using the Smoluchowski Coagulation Model 用Smoluchowski凝固模型研究铁蛋白自组装

IF 1.1

Biochemistry (Moscow), Supplement Series A: Membrane and Cell Biology Pub Date : 2025-04-24 DOI: 10.1134/S1990747824700442

M. S. Gette, D. M. Fedorov, O. M. Tilinova, Yu. L. Ryzhykau, A. V. Vlasov

{"title":"Studying Ferritin Self-Assembly Using the Smoluchowski Coagulation Model","authors":"M. S. Gette, D. M. Fedorov, O. M. Tilinova, Yu. L. Ryzhykau, A. V. Vlasov","doi":"10.1134/S1990747824700442","DOIUrl":"10.1134/S1990747824700442","url":null,"abstract":"Ferritin is an iron-storage protein complex performing vital functions in various living organisms. Its properties are significantly defined by a topology of the protein globule and the structural arrangement of subunits on its surface. Understanding the mechanisms of ferritin self-assembly could open up new ways of functionalization of its surface coating with a number of applications in biotechnology. In this study, we investigated the self-assembly of ferritin using the Smoluchowski coagulation model. We numerically solved the Smoluchowski differential equations for various models, involving the formation of hybrid ferritins with two types of subunits (for instance, H and L). Our model incorporates different reaction schemes and provides insights into the kinetics of oligomer assembly. The results reveal that our model can accurately describe the temporal dynamics of ferritin assembly, predicting the formation of intermediate states and fully assembled globules. The extension of the model using a four-dimensional coagulation kernel enables a detailed description of hybrid ferritin assembly, offering a new perspective on the complexity of self-assembly processes in heterooligomeric protein systems. Overall, our findings provide a robust framework for understanding the dynamics of ferritin assembly, offering insights that could be generalized to other carriers and help in designing more effective experimental approaches to study these mechanisms.","PeriodicalId":484,"journal":{"name":"Biochemistry (Moscow), Supplement Series A: Membrane and Cell Biology","volume":"19 1","pages":"46 - 51"},"PeriodicalIF":1.1,"publicationDate":"2025-04-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143865454","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}

引用次数: 0

Nanosystems Consisting of Iron Oxide and Serum Albumin As a Platform for Drug Delivery into Cells: Analysis of the Cy5-HSA@IONPs Nanosystem Components before and after Accumulation by Tumor Cells 由氧化铁和血清白蛋白组成的纳米系统作为药物递送到细胞的平台：Cy5-HSA@IONPs纳米系统成分在肿瘤细胞积累前后的分析

IF 1.1

Biochemistry (Moscow), Supplement Series A: Membrane and Cell Biology Pub Date : 2025-04-24 DOI: 10.1134/S1990747824700557

A. V. Toroptseva, A. A. Markova, M. T. Nguyen, M. I. Abdullina, M. V. Motyakin, O. A. Mayorova, A. V. Bychkova

{"title":"Nanosystems Consisting of Iron Oxide and Serum Albumin As a Platform for Drug Delivery into Cells: Analysis of the Cy5-HSA@IONPs Nanosystem Components before and after Accumulation by Tumor Cells","authors":"A. V. Toroptseva, A. A. Markova, M. T. Nguyen, M. I. Abdullina, M. V. Motyakin, O. A. Mayorova, A. V. Bychkova","doi":"10.1134/S1990747824700557","DOIUrl":"10.1134/S1990747824700557","url":null,"abstract":"The hybrid shell@core nanosystem Cy5-HSA@IONPs containing magnetic iron oxide core and Cy5-HSA shell was engineered and characterized by composition, stability and integrity using fluorescent measurements, UV/Vis-spectrophotometry (particularly, Bradford protein assay), dynamic light scattering, electron magnetic resonance in solution, in cells and cell lysates. Conjugation of HSA to Cyanine 5 (Cy5) fluorescent dye was carried out for qualitative and quantitative estimations. The research included incubation of hybrid nanosystems with human adenocarcinoma cells (HCT116 and MCF7, colon and breast cell lines, respectively) followed by cell lysis, magnetic separation and quantitative determination of protein on the surface of the magnetic cores after mild cell lysis and in suspension of cell residues. The presence of the albumin coating on the surface of magnetic cores (IONPs) upon delivery to the cells has been proven and the effectiveness of the engineered hybrid particles as a drug (photosensitizer) delivery platform has been discussed.","PeriodicalId":484,"journal":{"name":"Biochemistry (Moscow), Supplement Series A: Membrane and Cell Biology","volume":"19 1","pages":"122 - 126"},"PeriodicalIF":1.1,"publicationDate":"2025-04-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143865418","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}

引用次数: 0

Lysosome Functioning as a Process that Slows Down Aging 溶酶体在延缓衰老过程中的作用

IF 1.1

Biochemistry (Moscow), Supplement Series A: Membrane and Cell Biology Pub Date : 2025-04-24 DOI: 10.1134/S1990747824700387

V. A. Alekhin, S. F. Nazarova, D. R. Gizatullina, E. T. Lopatina, M. P. Kirpichnikov, M. A. Ostrovsky, V. I. Gordeliy, N. S. Ilyinsky

{"title":"Lysosome Functioning as a Process that Slows Down Aging","authors":"V. A. Alekhin, S. F. Nazarova, D. R. Gizatullina, E. T. Lopatina, M. P. Kirpichnikov, M. A. Ostrovsky, V. I. Gordeliy, N. S. Ilyinsky","doi":"10.1134/S1990747824700387","DOIUrl":"10.1134/S1990747824700387","url":null,"abstract":"Loss of macroautophagy and loss of proteostasis are hallmarks of aging. Lysosomal dysfunction is one of the reasons for the accumulation of non-degradable defective macromolecules, protein aggregates, and damaged organelles in the cells of organisms. The summary of experimental data obtained so far indicates an age-dependent weakening of lysosomal function. We hypothesize that lysosomes, as part of the autophagy and signaling network, are of great importance for longevity. In this review, we summarize information on the mechanisms of lysosomal influence on the lifespan and existing approaches to stimulate lysosomal function for longevity. We emphasize that factors such as intraluminal pH and interactions of lysosome-associated regulatory proteins (mTOR, AMPK, TFEB, and others) are related to aging. Stimulation of lysosomal function by dietary restriction, pharmacological or optogenetic approaches could be one of the promising interventions for anti-aging protection. In particular, a novel group of lysosomal optogenetic tools with high specificity of action could be a breakthrough in aging research.","PeriodicalId":484,"journal":{"name":"Biochemistry (Moscow), Supplement Series A: Membrane and Cell Biology","volume":"19 1","pages":"3 - 11"},"PeriodicalIF":1.1,"publicationDate":"2025-04-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143865379","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}

引用次数: 0

Searching for the Role of Mechanosensitive PIEZO1 Channel in Cell Motility and Migration of Human Aggressive Melanoma SK-MEL-2 Cells 机械敏感的PIEZO1通道在人类侵袭性黑色素瘤SK-MEL-2细胞运动和迁移中的作用

IF 1.1

Biochemistry (Moscow), Supplement Series A: Membrane and Cell Biology Pub Date : 2025-04-24 DOI: 10.1134/S1990747824700569

V. Y. Vasileva, V. I. Chubinskiy-Nadezhdin

{"title":"Searching for the Role of Mechanosensitive PIEZO1 Channel in Cell Motility and Migration of Human Aggressive Melanoma SK-MEL-2 Cells","authors":"V. Y. Vasileva, V. I. Chubinskiy-Nadezhdin","doi":"10.1134/S1990747824700569","DOIUrl":"10.1134/S1990747824700569","url":null,"abstract":"Melanoma is one of the most common cancers in the world, especially in young adults. Due to the emergence of the resistance for treatments, the identification of novel potential molecular targets is of specific importance. Previously, we have identified functional mechanosensitive Piezo1 channels in the plasma membrane of aggressive melanoma SK-MEL-2 cell line and evidenced that selective Piezo1 activation by chemical agonist Yoda1 completely prevented melanoma spheroid formation. Here, we expanded the number of experimental tests to reveal the role of Piezo1 activity in pathophysiological reactions of SK-MEL-2 cells that are linked with cell motility and migration. In particular, we assessed the melanoma cell motility using wound closure and spheroid reactivation assay in the presence of Piezo1-selective chemical activator, Yoda1. Ca2+ imaging allowed us to detect variations in a number of functionally active Piezo1 between the cells on the surface of melanoma spheroids. The processes of melanoma spheroid reactivation were unaffected by Yoda1. At the same time, wound healing assay showed a decrease of SK-MEL-2 migratory activity in the presence of Yoda1. Our results demonstrate differential roles of Piezo1 in regulation of migratory properties of melanoma cells.","PeriodicalId":484,"journal":{"name":"Biochemistry (Moscow), Supplement Series A: Membrane and Cell Biology","volume":"19 1","pages":"127 - 132"},"PeriodicalIF":1.1,"publicationDate":"2025-04-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143865380","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}

引用次数: 0

HSP70-NK and Target Cell Interactions: Implication to the Anticancer Effects HSP70-NK与靶细胞相互作用：对抗癌作用的影响

IF 1.1

Biochemistry (Moscow), Supplement Series A: Membrane and Cell Biology Pub Date : 2025-04-24 DOI: 10.1134/S1990747824700491

M. A. Shevchenko, O. V. Ovsyanikova, M. V. Grechikhina, D. G. Garbuz, A. A. Boyko, M. B. Evgen’ev, O. G. Zatsepina, E. I. Kovalenko, A. M. Sapozhnikov

{"title":"HSP70-NK and Target Cell Interactions: Implication to the Anticancer Effects","authors":"M. A. Shevchenko, O. V. Ovsyanikova, M. V. Grechikhina, D. G. Garbuz, A. A. Boyko, M. B. Evgen’ev, O. G. Zatsepina, E. I. Kovalenko, A. M. Sapozhnikov","doi":"10.1134/S1990747824700491","DOIUrl":"10.1134/S1990747824700491","url":null,"abstract":"Heat shock protein 70 (HSP70) and HSP70-based vaccine constructs are perspective tools for anticancer therapy. One of the established anticancer effects of HSP70 is activating natural killer (NK) cells. Moreover, HSP70-activated NK cells demonstrate effectiveness in preclinical glioblastoma and lung cancer models. It is thought that the expression of HSP70 on the surface of cancer cells stimulates the cytotoxic activity of NK cells. Regarding this, the spatial aspects of NK cells with cancer cells in the presence of HSP70 are essential but not properly investigated. Using confocal laser scanning microscopy, we have recently shown that exogenously added HSP70 is absorbed on the surface of dying NK cells, while live NK cells internalize the protein. In the present study, we visualized the spatial aspects of NK cell interactions with the target breast cancer cell line BT-20 in the presence of exogenous HSP70. We traced the uptake of HSP70 by BT-20 cells and investigated NK cell behavior and contacts with BT-20 cells in the presence of HSP70. The data obtained in the present study on the interaction of BT-20 cells, NK cells, and HSP70 revealed novel aspects of HSP70-mediated anticancer efficacy of NK cells.","PeriodicalId":484,"journal":{"name":"Biochemistry (Moscow), Supplement Series A: Membrane and Cell Biology","volume":"19 1","pages":"79 - 83"},"PeriodicalIF":1.1,"publicationDate":"2025-04-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143865416","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}

引用次数: 0

SPR Analysis of Adrenodoxin Interaction with CYP11B1 Embedded into the Lipid Bilayer 肾上腺素还毒素与嵌入脂质双分子层的CYP11B1相互作用的SPR分析

IF 1.1

Biochemistry (Moscow), Supplement Series A: Membrane and Cell Biology Pub Date : 2025-04-24 DOI: 10.1134/S1990747824700466

O. V. Gnedenko, E. O. Yablokov, L. A. Kaluzhskiy, Y. V. Mezentsev, T. V. Tsybruk, M. A. Karpova, A. S. Ivanov

{"title":"SPR Analysis of Adrenodoxin Interaction with CYP11B1 Embedded into the Lipid Bilayer","authors":"O. V. Gnedenko, E. O. Yablokov, L. A. Kaluzhskiy, Y. V. Mezentsev, T. V. Tsybruk, M. A. Karpova, A. S. Ivanov","doi":"10.1134/S1990747824700466","DOIUrl":"10.1134/S1990747824700466","url":null,"abstract":"Surface plasmon resonance (SPR) is a relevant method for studying interactions involving membrane proteins, mitochondrial proteins in particular. Currently, there is only a little information on the interaction of mitochondrial proteins with their partners, such as electron carriers, in lipid microenvironment. In this work, using cytochrome P450 11β-hydroxylase (CYP11B1) and adrenodoxin (Adx) as an example, we showed that SPR analysis can be used for studying protein–protein interactions in lipid microenvironment. Kinetic and equilibrium parameters of the CYP11B1-Adx complexes in lipid bilayers of different compositions were determined. Our results reveal that the CYP11B1-Adx complex in cardiolipin-containing bilayer demonstrates increased stability: nearly tenfold reduction in the equilibrium dissociation constant (Kd) compared to the bilayer without cardiolipin. The interaction between redox proteins is highly sensitive to the lipid structure. The immobilized protein and the lipid composition should be selected in each case, but overall, the obtained data are highly complementary for studying membrane proteins.","PeriodicalId":484,"journal":{"name":"Biochemistry (Moscow), Supplement Series A: Membrane and Cell Biology","volume":"19 1","pages":"60 - 65"},"PeriodicalIF":1.1,"publicationDate":"2025-04-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143865545","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}

引用次数: 0

Identification of Conventional Dendritic Cell Subpopulations in Conducting Airway Mucosa 导气管粘膜常规树突状细胞亚群的鉴定

IF 1.1

Biochemistry (Moscow), Supplement Series A: Membrane and Cell Biology Pub Date : 2025-04-24 DOI: 10.1134/S1990747824700508

M. A. Shevchenko, E. A. Servuli, A. K. Kryuchkova, I. S. Okhrimenko

{"title":"Identification of Conventional Dendritic Cell Subpopulations in Conducting Airway Mucosa","authors":"M. A. Shevchenko, E. A. Servuli, A. K. Kryuchkova, I. S. Okhrimenko","doi":"10.1134/S1990747824700508","DOIUrl":"10.1134/S1990747824700508","url":null,"abstract":"Dendritic cells belong to a heterogenic population of innate immune cells distributed across the tissues. These cells are considered to be potential therapeutic targets due to their ability to sense exogenous antigens and self-tissue damage. In the lungs, two subpopulations of conventional dendritic cells, cDC1 and cDC2, were identified. Distinguishing the populations is commonly performed by flow cytometry based on the different expressions of transcription factors and surface markers. Thus, after the exclusion of precursors and the cells rather than dendritic cells, G protein-coupled receptor XCR1 and membrane glycoprotein SIRPA (CD172a) are used to characterize cDC1 as XCR1hiCD172alo and cDC2 as XCR1loCD172ahi. Analysis by flow cytometry permits the accurate identification of the subpopulations; however, to define the precise location of cDC1 and cDC2 in relation to the structural tissue cells, confocal laser scanning microscopy (CLSM) is required. Together with the advantage of spatial cell distribution identification, CLSM has two limitations compared to flow cytometry. The former is the limited number of antibodies that can be simultaneously applied. Besides, not all the antibodies suitable for cell staining and flow cytometry show stable antigen recognition during the tissue staining and CLSM. In this study, we applied anti-XCR1 and anti-CD172a to visualize cDC1 and cDC2 in conducting airway mucosa of mice. We detect the distribution of these cells in a steady state and upon the airway inflammation.","PeriodicalId":484,"journal":{"name":"Biochemistry (Moscow), Supplement Series A: Membrane and Cell Biology","volume":"19 1","pages":"84 - 88"},"PeriodicalIF":1.1,"publicationDate":"2025-04-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143865417","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}

引用次数: 0

Expression and Purification of an Archaeal Cysteine-less LOV Domain 古菌无半胱氨酸 LOV 结构域的表达与纯化

IF 1.1

Biochemistry (Moscow), Supplement Series A: Membrane and Cell Biology Pub Date : 2025-04-24 DOI: 10.1134/S199074782470048X

I. I. Natarov, O. Y. Semenov, A. A. Remeeva, I. Y. Gushchin

{"title":"Expression and Purification of an Archaeal Cysteine-less LOV Domain","authors":"I. I. Natarov, O. Y. Semenov, A. A. Remeeva, I. Y. Gushchin","doi":"10.1134/S199074782470048X","DOIUrl":"10.1134/S199074782470048X","url":null,"abstract":"LOV (Light Oxygen Voltage) domains are ubiquitous blue light sensors widespread in all three domains of life. They mediate signal transduction in diverse biological processes and can be useful for practical applications in optogenetics and fluorescence microscopy. The canonical mechanism of LOV domains signal transduction is based on formation of the covalent adduct between flavin chromophore and the highly conserved cysteine in the active site of the protein leading to downstream signaling cascade. However, previous studies showed that there are natural cysteine-less LOV domains enabling signal transduction via flavin photoreduction. In the absence of the conserved cysteine, photoexcitation causes flavin one electron reduction to the neutral semiquinone radical state and subsequent structural reorganizations as in the canonical photocycle. In this work, previously uncharacterized natural cysteine-less LOV domain from thermophilic haloarchaeon Halanaeroarchaeum sulfurireducens, named HsuLOV, was successfully expressed in Escherichia coli and purified in soluble form. Although HsuLOV bound a small amount of flavin when overexpressed in E. coli, the chromophore load was increased by protein incubation with flavin mononucleotide in vitro. It was demonstrated that fluorescence spectra of purified and chromophore-loaded HsuLOV are typical for LOV domains. Finally, the comparison of absorption spectra of dark- and light-adapted states of HsuLOV revealed flavin photoreduction to the neutral semiquinone radical state, thus confirming photoactivity of HsuLOV.","PeriodicalId":484,"journal":{"name":"Biochemistry (Moscow), Supplement Series A: Membrane and Cell Biology","volume":"19 1","pages":"73 - 78"},"PeriodicalIF":1.1,"publicationDate":"2025-04-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143865415","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}

引用次数: 0