Russian Journal of Bioorganic Chemistry最新文献

{"title":"Characterization of Combined Effects of Reactive Oxygen Metabolites, Complement System, and Antimicrobial Peptides In Vitro","authors":"I. A. Krenev, E. V. Egorova, N. P. Gorbunov, V. A. Kostevich, A. V. Sokolov, A. S. Komlev, Y. A. Zabrodskaya, O. V. Shamova, M. N. Berlov","doi":"10.1134/S1068162025010996","DOIUrl":"10.1134/S1068162025010996","url":null,"abstract":"<p><b>Objective:</b> Phagocytes activation results in the production of reactive oxygen metabolites exerting antimicrobial and host-damaging activity. Although the main pool of papers shows their potentiating action on a key humoral nexus of innate immunity, complement system, the data are controversial. Combined action of reac­tive oxygen metabolites with antimicrobial peptides of phagocytes also remains poorly characterized. <b>Methods:</b> We have investigated the influence of oxidative burst products on complement activation in various <i>in vitro</i> models and assessed the combined bactericidal action of hypochlorous acid with antimicrobial peptides. <b>Results and Discussion:</b> Hydrogen peroxide, including that in medium with Fe-EDTA did not affect parameters of complement activity in human blood serum. HOCl in millimolar concentrations stimulated production of C3a and C5a anaphylatoxins in 80% serum, the effect was inhibited by EDTA. We have identified bivalent ions-independent C5 cleavage in the presence of 16 mM HOCl. At the same time, HOCl served as an inhibitor of the alternative complement pathway in the model of membrane-associated activation in 5% serum in the presence of Mg-EGTA and rabbit erythrocytes. It inhibited production of C3a (IC<sub>50</sub> ~4 mM) and C5a as well as serum hemolytic activity (IC<sub>50</sub> ~0.2 mM). The inhibition of C5a generation was less pro­nounced in the presence of relatively high HOCl concentration (4–16 mM). Decrease in anaphylatoxins generation was also observed in the system with zymosan in 5% serum with Mg-EGTA. Under similar conditions but without activating surfaces, moderate HOCl concentrations enhanced C3a accumulation and C5a accumulation; EDTA inhibited this effect completely (C3a) or partially (C5a). Finally, in 70% serum, 16 mM HOCl enhanced the anaphylatoxins accumulation in the absence of zymosan but it inhibited this process almost completely under the conditions of the zymosan-triggered amplification loop. According to our hypothesis, HOCl can attack the thioester bond in C3 protein to form C3(HOCl) adduct which is capable of fluid-phase C3 and C5 convertases formation; however, the attack of the same group in C3b can prevent its covalent fixation on membranes and blocks the complement amplification loop. In addition, the transformation of C3 to C3(HOCl) which is not able to serve as a substrate of C3 convertases may also be responsible for complement inhibition. Besides, we have demonstrated the additive character of the combined action of HOCl with antimicrobial peptides (LL-37 cathelici­din and α-defensins HNPs) against <i>Listeria monocytogenes</i> and <i>Escherichia coli</i>. <b>Conclusions:</b> According to our results, hydrogen peroxide and hydroxyl radical do not participate in complement modulation. HOCl is an activator of fluid-phase and an inhibitor of surface complement activation. HOCl may also induce complement-independent C5 cleavage in serum. HOCl and antimicrobi","PeriodicalId":758,"journal":{"name":"Russian Journal of Bioorganic Chemistry","volume":"51 1","pages":"235 - 250"},"PeriodicalIF":1.1,"publicationDate":"2025-02-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143396446","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Exploring the Anticancer Properties of β-Sitosterol","authors":"Sainath Bhavsar,&nbsp;Sunita Pachori,&nbsp;Jitendra Patil","doi":"10.1134/S106816202501008X","DOIUrl":"10.1134/S106816202501008X","url":null,"abstract":"<p>Cancer is considered one of the most feared and dreaded diseases. According to the World Health Organization, it is the second leading cause of death worldwide. The uncontrollable division of cells is the main reason for cancer. Research is still ways to cure this hreatening disease, but very few significant results have been obtained to date. β-Sitosterol resembles the cholesterol moiety and belongs to the class of phytosterols. It is a white, waxy powdered sterol. It has been found that this compound inhibits the growth of cancerous cells. This review is devoted to the anticancer properties of the compound β-sitosterol on cancerous cells.</p>","PeriodicalId":758,"journal":{"name":"Russian Journal of Bioorganic Chemistry","volume":"51 1","pages":"171 - 176"},"PeriodicalIF":1.1,"publicationDate":"2025-02-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143396447","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Spectral-Luminescent Properties of Products of Interaction of Polyfluorinated Pyrazoline-Containing Pyrylium Dyes with Bovine Serum Albumin and Amino Acids","authors":"V. V. Shelkovnikov,&nbsp;D. D. Doroshenko,&nbsp;I. Yu. Kargapolova,&nbsp;P. A. Kozlakov,&nbsp;N. A. Shestakov,&nbsp;Yu. S. Sotnikova","doi":"10.1134/S1068162025010200","DOIUrl":"10.1134/S1068162025010200","url":null,"abstract":"<p><b>Objective:</b> Studies of the reaction ability of fluorescent dyes to produce labelled proteins and amino acids are important for bioengineering and biomedicine, particularly for cell visualization by bioimaging and the study of the structure of labelled proteins by biophysical methods. Pyrylium dyes can interact with the amino groups of proteins to form luminescent products, which allows them to be used in the field of proteomics. It is interesting to study the pyrylium conjugates that contain both a polyfluorinated fragment responsible for increased lipophilicity of the protein conjugates and a pyrazoline fragment responsible for anticancer activity. <b>Methods:</b> The pyrylium dyes that contain the pyrazoline fragment and dialkylamino substituents (piperidino-, dibutylamino-, and 4-hydroxypiperidino-) in the donor part of the polyfluorinated aromatic ring have been synthesized by the Knoevenagel condensation reaction. The reaction of pyrylium dyes with compounds containing the primary amino group has been performed to obtain the pyridinium dyes by the ANRORC (Addition of Nucleophiles, Ring Opening and Ring Closure) Mechanism.<b> Results and Discussion:</b> The ability of the pyrуlium dyes to react with bovine serum albumin (BSA) and amino acids, such as Lys, Arg, Cys, and Phe to form pyridinium luminophore has been demonstrated. The spectral-luminescent properties of the resulting luminophores have been investigated. The product of the reaction of the pyrуlium dye (<i>Е</i>)-2,6-dimethyl-4-(4-{3-phenyl-5-[2,3,5,6-tetrafluorо-4-(piperidine-1-yl)phenyl]-4,5-dihydro-1<i>Н</i>-pуrazol-1-yl}-styryl)pyrylium tetrafluoroborate with Lys has been isolated, and its structure has been confirmed by NMR spectroscopy. The binding site of the pyrylium dyes with BSA, i.e., the ε-amino group of Lys, has been determined. Together with the pyridinium luminophores, hydrolysis products are formed in aqueous solutions, which are not bound to the protein and absorb in the short wavelength region. The amount of the pyrylium dye bound to BSA has been calculated to be 2 : 1.The synthesized pyrylium dyes react with BSA in the phosphate buffer-methanol mixture (pH 7.4) 3–4 orders of magnitude faster than the well-known julolidine dye Py-1. The relative reaction rates of (<i>Е</i>)-2,6-dimethyl-4-(4-{3-phenyl-5-[2,3,5,6-tetrafluorо-4-(4-hydroxypiperidine-1-yl)phenyl]-4,5-dihydro-1<i>Н</i>-pуrazol-1-yl}styryl) pyrуlium tetrafluoroborate with amino acids have been evaluated to be as follows: Lys &gt; Cys &gt;&gt; Phe ≥ Arg. <b>Conclusions:</b> The synthesized polyfluoro pyrylium-pyrazoline dyes have the application perspective in the field of bioimaging, proteomics, and biomedicine due to the high conjugation rate and efficiency with BSA and amino acids.</p>","PeriodicalId":758,"journal":{"name":"Russian Journal of Bioorganic Chemistry","volume":"51 1","pages":"216 - 228"},"PeriodicalIF":1.1,"publicationDate":"2025-02-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143396578","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Visualization of H3K9me3 in Embryoid Bodies Using Genetically Encoded Fluorescent Sensor MPP8-Green","authors":"A. I. Stepanov,&nbsp;E. B. Zhigmitova,&nbsp;E. B. Dashinimaev,&nbsp;A. A. Galiakberova,&nbsp;L. V. Putlyaeva,&nbsp;K. A. Lukyanov,&nbsp;N. G. Gurskaya","doi":"10.1134/S1068162025010236","DOIUrl":"10.1134/S1068162025010236","url":null,"abstract":"<p><b>Objective:</b> Histone modifications play a crucial role in shaping the epigenetic landscape of chromatin, influencing its structure and function. These histone modifications change throughout the cell cycle and during processes such as morphogenesis, differentiation, stress adaptation, and aging. Among these, histone H3 Lys9 trimethylation (H3K9me3) plays a critical role in gene silencing and cellular identity. Despite the importance of histone modifications in regulating gene expression, the dynamic visualization of these modifications in live cells remains a significant challenge. Here, we aim to develop a method to track H3K9me3 changes of induced pluripotent stem cells (iPSCs) during spontaneous differentiation into embryoid bodies (EBs) with the genetically encoded fluorescent sensor. <b>Methods:</b> We created a stable iPSC line, KUIFMSi004-A-1, expressing the MPP8-Green fluorescent sensor, which binds H3K9me3. This sensor combines two copies of the natural M-phase phosphoprotein 8 (MPP8) reader domain with the green fluorescent protein mNeonGreen. The iPSC line was induced to form EBs, and the distribution of H3K9me3 was monitored using live-cell fluorescence microscopy. <b>Results and Discussion:</b> The expression of the MPP8-Green sensor allowed real-time visualization of H3K9me3 modifications during spontaneous differentiation of iPSCs into EBs. We observed two distinct groups of cells with different patterns of H3K9me3 distribution: one with characteristic chromatin “dots” and another with diffuse sensor distribution. The sensor enabled tracking of epigenetic landscape changes during differentiation, providing insights into the dynamics of H3K9me3 during early embryoid body formation. <b>Conclusions:</b> Our study demonstrates the potential of using the MPP8-Green sensor to monitor the dynamic changes of H3K9me3 during iPSC differentiation. This method offers a novel approach for studying the temporal and spatial regulation of histone modifications in live cells, advancing our understanding of epigenetic regulation during development.</p>","PeriodicalId":758,"journal":{"name":"Russian Journal of Bioorganic Chemistry","volume":"51 1","pages":"229 - 234"},"PeriodicalIF":1.1,"publicationDate":"2025-02-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143396581","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Recent Advances in Liposomes Efficacy as a Drug Delivery Platform for Melanoma Treatment","authors":"Elham Zarenezhad,&nbsp;Zahra Kazeminejad,&nbsp;Mahsa Rostami Chijan,&nbsp;Mahmoud Osanloo,&nbsp;Ensieh Nournia,&nbsp;Abdolmajid Ghasemian,&nbsp;Mahrokh Marzi","doi":"10.1134/S1068162025010030","DOIUrl":"10.1134/S1068162025010030","url":null,"abstract":"<p>Melanoma is the predominant skin cancer with the potential for metastasis. The disease has inferred an enhancing trend worldwide during recent years, needing proper therapeutic approaches. Liposomes place among the leading vesicular drug delivery platforms, permitting controlled release, targeted therapy, and hence exerting minimal off-target toxicity. They can be prepared in various sizes, entrap various hydrophilic and hydrophobic medicines, and load various lipid- and water -soluble drugs and compounds in relatively high amounts. Liposomes protect the cargo and endure within the biological structures against degradation and clearance. They are biologically inert and biodegradable and decrease the toxicity, antigenicity, or pyrogenicity of medicinal compounds. The combination therapy of liposomal delivery with chemotherapeutics and immunotherapy approaches has been promising. In this study, we provide the results of a biological activity evaluation of liposomes in melanoma cancer. This review will be useful in different disciplines, such as organic chemistry, pharmacology, medicinal chemistry, biochemistry, and oncology.</p>","PeriodicalId":758,"journal":{"name":"Russian Journal of Bioorganic Chemistry","volume":"51 1","pages":"93 - 116"},"PeriodicalIF":1.1,"publicationDate":"2025-02-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143396674","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Chemotherapeutic Boron-Containing Homocysteinamides of Human Serum Albumin","authors":"M. Wang,&nbsp;S. A. Tsyrempilov,&nbsp;I. A. Moskalev,&nbsp;O. D. Zakharova,&nbsp;A. I. Kasatova,&nbsp;V. N. Sil’nikov,&nbsp;T. S. Godovikova,&nbsp;T. V. Popova","doi":"10.1134/S1068162025010297","DOIUrl":"10.1134/S1068162025010297","url":null,"abstract":"<p><b>Objective:</b> A combination of boron neutron capture therapy and chemotherapy can ensure good efficacy in cancer treatment. The development of therapeutic constructs that combine these two functions, specifically the possibility of <i>in vitro</i> and <i>in vivo</i> visualization and a convenient platform for selective delivery to the tumor, is of great relevance today. <b>Methods:</b> In this study, we focused on human serum albumin, a well-known drug delivery platform, and developed, based on albumin functionalized with boron clusters, therapeutic constructs, which as analogs of the chemotherapeutic molecule gemcitabine and signaling molecules. To create such constructs, we developed new analogs of homocysteine thiolactone containing <i>closo</i>-dodecaborate or cobalt bis(dicarbollide) and a gemcitabine analog containing <i>closo</i>-dodecaborate attached to the C⁵ carbon atom of the nitrogenous base. <b>Results and Discussion:</b> It was demonstrated that the the conjugates modified with the gemcitabine analogs exhibit increased cytotoxicity against human glioblastoma cell lines. Among the obtained conjugates, the highest cytotoxicity is demonstrated by that containing cobalt bis(dicarbollide). The resulting structures accumulate well in the cytoplasm of cancer cells. The albumin conjugate containing cobalt <i>bis</i>(dicarbollide) and the boron-containing gemcitabine analog is capable of accumulating in the nuclei of T98G cell lines. <b>Conclusions:</b> The resulting albumin constructs showed sufficient <i>in vitro</i> activity against human glioma cells. It is expected that the obtained therapeutic conjugates will significantly enhance the anticancer efficacy of irradiation with epithermal neutrons. Combining a chemotherapeutic residue and a boron-containing group in a single construct provides potential for more effective glioma therapy.</p>","PeriodicalId":758,"journal":{"name":"Russian Journal of Bioorganic Chemistry","volume":"51 1","pages":"354 - 371"},"PeriodicalIF":1.1,"publicationDate":"2025-02-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143396453","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Preparation of Boron-Containing S-Nitrosothiol Based on Homocysteinylamides of Human Serum Albumin for Combined NO-Chemical and Boron-Neutron-Capture Therapy","authors":"T. V. Popova,&nbsp;M. Van,&nbsp;T. N. Kurochkin,&nbsp;S. A. Tsyrempilov,&nbsp;O. D. Zakharova,&nbsp;V. N. Silnikov,&nbsp;T. S. Godovikova","doi":"10.1134/S1068162025010194","DOIUrl":"10.1134/S1068162025010194","url":null,"abstract":"<p><b>Objective:</b> The strategic aim of this work is to create a fluorophore-labelled, clinically relevant exogenous NO donor carrying a boron-containing compound residue based on human serum albumin (HSA) for the implementation of combined NO-chemotherapy and boron-neutron-capture therapy. <b>Methods:</b> By selective modification of the Cys34 residue of albumin with a maleimide derivative of a fluorescent dye and subsequent N-homocysteinylation with a thiolactone derivative of homocysteine containing a clozo-dodecaborate residue, a nanoconstruct for boron-neutron-capture therapy was obtained. An analogue based on the natural modifier, boron-containing homocysteine thiolactone, was synthesised by alkylation of the amino group of thiolactone with a dioxonium derivative of clozo-dodecaborate. Post-synthetic modification of the lysine residues of the protein using the boron thiolactone of homocysteine provided the introduction of SH groups into the protein and the possibility of subsequent trans-S-nitrosylation of the protein using S-nitrosoglutathione. <b>Results and Discussion:</b> It was found that 2 M of NO was conjugated to 1 M of boron-containing HSA. Boron-containing S-nitrosothiol based on albumin homocysteinylamide, without epithermal neutron irradiation, was demonstrated to be more cytotoxic against human glioblastoma cell lines than the boron-containing albumin conjugate. <b>Conclusions:</b> Thus, the approach used allows obtaining a boron-enriched structure based on a biocompatible tumor-specific protein, containing a fluorescent label and an increased number of S-nitroso groups. It is necessary for the manifestation of a chemotherapeutic effect of the construct. The practical significance of this structure lies in the possibility of a cancer treatment, combining chemo- and boron-neutron capture therapy.</p>","PeriodicalId":758,"journal":{"name":"Russian Journal of Bioorganic Chemistry","volume":"51 1","pages":"202 - 215"},"PeriodicalIF":1.1,"publicationDate":"2025-02-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143396577","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Live-Cell Visualization of Histone Modification Using Bimolecular Complementation","authors":"A. I. Stepanov,&nbsp;L. V. Putlyaeva,&nbsp;A. A. Shuvaeva,&nbsp;M. A. Andrushkin,&nbsp;M. S. Baranov,&nbsp;N. G. Gurskaya,&nbsp;K. A. Lukyanov","doi":"10.1134/S1068162025010261","DOIUrl":"10.1134/S1068162025010261","url":null,"abstract":"<p><b>Objective:</b> The study of histone post-translational modifications (PTMs) is a rapidly developing field, yet the tools available for detecting and interpreting these modifications are limited. Histone modifications, such as methylation, acetylation, and phosphorylation, play a crucial role in regulating chromatin dynamics and gene expression. Specific binding of histone modification “reader” domains (HMRDs) is central to this regulation, allowing for the recruitment of proteins that facilitate chromatin remodeling. This research aims to develop genetically encoded sensors based on HMRDs to study histone modifications in live cells, offering a more efficient and flexible method for studying epigenetic changes. <b>Methods:</b> We designed genetically encoded sensors that utilize HMRDs and splitFAST to bind specifically to different histone modifications. These sensors were incorporated into cells to track the dynamic changes in histone modifications. The performance of these sensors was evaluated through live-cell imaging, using fluorescent microscopy to monitor histone modifications. <b>Results and Discussion:</b> The genetically encoded sensors demonstrated high specificity and sensitivity to various histone modifications. Sensors based on SplitFAST and HMRDs MPP8 and AF9 exhibited specific distributions for H3K9me3 and H3K9ac. Moreover, the combination of these two domains with different parts of SplitFAST showed spatial proximity between H3K9me3 and H3K9ac. These findings suggest that the integration of HMRD-based sensors, MPP8 and AF9, with SplitFAST could provide valuable tools for live-cell monitoring of histone modifications and their roles in gene regulation and cellular response mechanisms. <b>Conclusions:</b> The development of genetically encoded sensors for histone modifications based on HMRDs provides a powerful new tool for studying chromatin dynamics in live cells. These sensors offer a more direct and real-time approach to understanding the complex mechanisms of histone modification and their impact on gene expression.</p>","PeriodicalId":758,"journal":{"name":"Russian Journal of Bioorganic Chemistry","volume":"51 1","pages":"320 - 329"},"PeriodicalIF":1.1,"publicationDate":"2025-02-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143396619","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"DHPMs C5 Amide Bearing Benzothiazole Moiety Inhibit AChE: Design, Synthesis, In Vitro and In Silico Studies","authors":"Pardis Samiei,&nbsp;Dara Dastan,&nbsp;Ahmad Ebadi","doi":"10.1134/S1068162025010285","DOIUrl":"10.1134/S1068162025010285","url":null,"abstract":"<p><b>Objective:</b> The well-accepted hypothesis in Alzheimer’s disease (AD) emphasizes the role of the cholinergic system in the disease. Therefore, the design and development of selective AChE inhibitors have been considered a promising strategy for AD treatment. Dihydropyrimidin-2-one is a privileged heterocyclic scaffold with a wide range of biological activities. <b>Methods:</b> Our initial docking studies indicated that substitution at the para position of the C4 aryl in the DHPM ring could interact with the gorge of the active site. Accordingly, we designed and synthesized eight DHPM derivatives to test the hypothesis. <b>Results and Discussion:</b> The results indicated that the DHPM with a benzothiazolyl carbamyl moiety at C5 and an OCF₃ group at the para position of the C4 aryl ring was the most potent inhibitor. The propargyloxy group was less potent than the OCF₃ group but had the same ligand efficiency. <b>Conclusions:</b> Introducing electron-rich substitutions of proper size increased activity, but the ligand efficiency remained constant. <i>In silico</i> studies revealed that the para substitution of the aryl ring interacted with the gorge residues in the R-enantiomer.</p>","PeriodicalId":758,"journal":{"name":"Russian Journal of Bioorganic Chemistry","volume":"51 1","pages":"340 - 353"},"PeriodicalIF":1.1,"publicationDate":"2025-02-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143396646","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Construct Design, Isolation, and Purification of the Human GPR17 Receptor Monomeric Form for Structural and Functional Studies","authors":"N. A. Safronova,&nbsp;A. P. Luginina,&nbsp;A. A. Sadova,&nbsp;M. B. Shevtsov,&nbsp;O. V. Moiseeva,&nbsp;V. I. Borshchevskiy,&nbsp;A. V. Mishin","doi":"10.1134/S106816202501025X","DOIUrl":"10.1134/S106816202501025X","url":null,"abstract":"<p><b>Objective:</b> G protein-coupled receptors (GPCRs) are a family of seven transmembrane domain proteins with more than 800 members in the human genome. They play a key role in the regulation of most of the processes in the human body and are the targets of one third of all modern drugs. Despite their importance in pharmacology, many GPCRs remain orphan receptors, i.e., their endogenous ligands are unknown. An orphan receptor GPR17, a class A GPCR representative, is predominantly expressed in the central nervous system and plays an important role in regulating the formation of the myelin sheath of neurons. It is a potential target for the development of new drugs against various disorders, such as multiple sclerosis, Alzheimer’s disease, and ischemia. The aim of this work was to prepare GPR17 for structural and functional studies starting from protein modification and ending with the receptor production. <b>Methods:</b> A screening of different genetically engineered constructs was performed, a series of point mutations were analyzed, and a significant number of potential ligands for this receptor were tested. The constructs were expressed in the Sf9 insect cell line using the Bac-to-Bac approach. The membrane fraction was extracted, the protein was solubilized into DDM/CHS detergent micelles, followed by purification by a metal affinity chromatography. Functional analysis of the purified protein included analytical gel filtration, polyacrylamide gel electrophoresis, and thermal stability analysis. <b>Results and Discussion:</b> Stabilizing point mutations were identified and the optimal position of the partner protein was found. The conditions for the expression, isolation, and purification of GPR17 were optimized to produce a sufficiently stable and monomeric protein sample suitable for further structural and functional studies. <b>Conclusions:</b> GPR17 is an orphan GPCR and the actively studied pharmacological target. The structural and functional investigation of this protein is a relevant problem for modern science and biomedicine. As a result of an in-depth study of GPR17, several stabilizing receptor modifications were developed, and optimal conditions for recombinant protein production were found. Our results can be used for further structural and functional studies of GPR17 and can serve as an example of a strategy that can be applied to perform investigation of other GPCRs.</p>","PeriodicalId":758,"journal":{"name":"Russian Journal of Bioorganic Chemistry","volume":"51 1","pages":"308 - 319"},"PeriodicalIF":1.1,"publicationDate":"2025-02-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143396616","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}