Biochimie最新文献_第3页

Development and evaluation of a new affiprobe that targets HER 3 positive cells using protein engineering 利用蛋白工程技术开发和评价一种靶向HER 3阳性细胞的新亲和物。

IF 3.3 3区生物学

Biochimie Pub Date : 2025-05-23 DOI: 10.1016/j.biochi.2025.05.008

Farnaz Karbasi , Rahman Emamzadeh , Mahboobeh Nazari

引用次数: 0

Structural and kinetic profiling of Leishmania braziliensis trypanothione reductase: A molecular model for the development of targeted therapies 巴西利什曼原虫锥虫硫酮还原酶的结构和动力学分析：开发靶向治疗的分子模型。

IF 3.3 3区生物学

Biochimie Pub Date : 2025-05-17 DOI: 10.1016/j.biochi.2025.05.006

Ana F. Gómez Garay , Jorge J. Alfonso , Aleff F. Francisco , Erika C.S. Araújo , Marcos R.M. Fontes , Carlos A.H. Fernandes , Soraya S. Pereira , Anderson M. Kayano , Leonardo A. Calderon , Jamile M. Macedo , Mateus F. Souza , Rudson J. Holanda , Juliana C. Sobrinho , Andreimar M. Soares

{"title":"Structural and kinetic profiling of Leishmania braziliensis trypanothione reductase: A molecular model for the development of targeted therapies","authors":"Ana F. Gómez Garay , Jorge J. Alfonso , Aleff F. Francisco , Erika C.S. Araújo , Marcos R.M. Fontes , Carlos A.H. Fernandes , Soraya S. Pereira , Anderson M. Kayano , Leonardo A. Calderon , Jamile M. Macedo , Mateus F. Souza , Rudson J. Holanda , Juliana C. Sobrinho , Andreimar M. Soares","doi":"10.1016/j.biochi.2025.05.006","DOIUrl":"10.1016/j.biochi.2025.05.006","url":null,"abstract":"<div><div>This study explores the recombinant protein expression, purification, and characterization of <em>Leishmania braziliensis</em> Trypanothione Reductase (<em>Lb</em>TR), an essential enzyme implicated in cutaneous leishmaniasis. Using <em>E. coli</em> as the host organism, the synthetic gene encoding <em>Lb</em>TR was successfully expressed and subsequently purified using Immobilized Metal Affinity Chromatography (IMAC) yielding 20 mg/L of highly pure <em>Lb</em>TR, verified by SDS-PAGE and isoelectric focusing. Comprehensive biochemical analyses were conducted to determine the recombinant enzyme's kinetic properties and structural features. Enzymatic assays revealed that <em>Lb</em>TR efficiently reduces its natural substrate, following Michaelis-Menten kinetics. Structural characterization, including dynamic light scattering and fluorescence spectroscopy, confirmed the protein's stability and homogeneity in solution under varying temperatures. Circular dichroism analysis corroborated the presence of significant α-helical and β-sheet content, aligning with the structural model generated with AlphaFold. Molecular dynamics simulations over 1 μs were employed to investigate the conformational dynamics of <em>Lb</em>TR in its native homodimeric form, complexed with essential cofactors and substrates. The results from these simulations offer valuable insights into the enzyme's structural behavior and catalytic mechanism, underscoring its potential as a target for therapeutic development against leishmaniasis.</div></div>","PeriodicalId":251,"journal":{"name":"Biochimie","volume":"235 ","pages":"Pages 14-28"},"PeriodicalIF":3.3,"publicationDate":"2025-05-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144103284","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Bothropstoxins I and II as potent phospholipase A2 molecules from Bothrops jararacussu to impair hepatitis C virus infection Bothropstoxins I和II是抑制丙型肝炎病毒感染的有效磷脂酶A2分子。

IF 3.3 3区生物学

Biochimie Pub Date : 2025-04-25 DOI: 10.1016/j.biochi.2025.04.006

Carina Machado Pereira , Jacqueline Farinha Shimizu , Natasha Marques Cassani , Igor Andrade Santos , Cintia Bittar , Adélia Cristina Oliveira Cintra , Suely Vilela Sampaio , Mark Harris , Paula Rahal , Ana Carolina Gomes Jardim

{"title":"Bothropstoxins I and II as potent phospholipase A2 molecules from Bothrops jararacussu to impair hepatitis C virus infection","authors":"Carina Machado Pereira , Jacqueline Farinha Shimizu , Natasha Marques Cassani , Igor Andrade Santos , Cintia Bittar , Adélia Cristina Oliveira Cintra , Suely Vilela Sampaio , Mark Harris , Paula Rahal , Ana Carolina Gomes Jardim","doi":"10.1016/j.biochi.2025.04.006","DOIUrl":"10.1016/j.biochi.2025.04.006","url":null,"abstract":"<div><div>Hepatitis C virus (HCV) (now classified <em>Hepacivirus hominis</em>) that infects an estimated 50 million individuals worldwide and causes chronic liver disease. The current treatment for infected patients primarily relies on direct-acting antivirals (DAAs). However, this treatment is marked by its high cost, numerous side effects, and documented instances of antiviral resistance. These challenges underscore the imperative for developing novel therapeutic strategies. In this framework, naturally occurring compounds have exhibited considerable medical significance attributable to their biological functionalities. Compounds extracted from snake venoms have evidenced antiviral efficacy against a variety of viral pathogens including <em>Orthoflavivirus denguei</em> (DENV), <em>Orthoflavivirus flavi</em> (YFV), <em>Orthoflavivirus zikaense</em> (ZIKV), and HCV. Here, the activity of 10 proteins isolated from snakes’ venom of <em>Bothrops</em> genus were evaluated against HCV replicative cycle. JFH-1 HCV system infected Huh-7.5 cell. Cell viability was measured simultaneously through MTT assay. Eight compounds inhibited up to 99 % of HCV infection, with the most potent inhibitory rates observed in BthTX-I and BthTX-II. These exhibited an SI of > 50 and 16,220, respectively, being able to block 84.7 % and 96 % of HCV infectivity. BthTX-II also demonstrated a protective effect in cells treated prior to HCV infection of approximately 86.7 %. Molecular docking calculations suggest interactions between the two proteins with HCV E1E2 glycoprotein complex. BthTX-II exhibited stronger interactions, indicated by 22 hydrophobic interactions. In conclusion, these compounds were shown to inhibit HCV infectivity by either acting on the virus particles or protecting the cells against infection.</div></div>","PeriodicalId":251,"journal":{"name":"Biochimie","volume":"235 ","pages":"Pages 39-48"},"PeriodicalIF":3.3,"publicationDate":"2025-04-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144002063","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Exploring the role of G-quadruplex DNA, and their structural polymorphism, in targeting small molecules for the design of anticancer therapeutics: Progress, challenges, and future directions 探索g -四重体DNA及其结构多态性在靶向小分子抗癌药物设计中的作用：进展、挑战和未来方向

IF 3.3 3区生物学

Biochimie Pub Date : 2025-04-17 DOI: 10.1016/j.biochi.2025.04.004

Soma Roy , Pulakesh Pramanik , Santanu Bhattacharya

引用次数: 0

Human prostacyclin and thromboxane synthases: Molecular interactions, regulation, and pharmacology 人前列环素和凝血素合成酶：分子相互作用、调控和药理学

IF 3.3 3区生物学

Biochimie Pub Date : 2025-04-11 DOI: 10.1016/j.biochi.2025.04.003

Pavel V. Ershov, Evgeniy O. Yablokov, Yuri V. Mezentsev, Alexis S. Ivanov

{"title":"Human prostacyclin and thromboxane synthases: Molecular interactions, regulation, and pharmacology","authors":"Pavel V. Ershov, Evgeniy O. Yablokov, Yuri V. Mezentsev, Alexis S. Ivanov","doi":"10.1016/j.biochi.2025.04.003","DOIUrl":"10.1016/j.biochi.2025.04.003","url":null,"abstract":"<div><div>Prostanoids are lipid mediators of the human body that are involved in the inflammation and platelet aggregation. Prostacyclin is a vasodilator and inhibitor of platelet aggregation, and a product of the enzymatic reaction catalyzed by prostacyclin synthase (PTGIS). Thromboxane is a vasoconstrictor and synthesized by thromboxane synthase (TBXAS1). An imbalance of prostanoids can accompany cardio-/cerebrovascular diseases and cancers. PTGIS and TBXAS1 are clinically relevant membrane-bound enzymes of the multigene family of cytochromes P450 (CYPs), also known as CYP8A1 and CYP5A1, respectively. Particular studies of these functional antagonists will contribute to the elucidation of pathogenic mechanisms. The purpose of this work was to analyze the literature landscape over a period of 2020–2024 in the field of biological, pharmacogenomic, and pharmacological features of PTGIS and TBXAS1 as well as to explore the potential of their regulation at the post-transcriptional and post-translational levels using systems biological analysis. The review discusses recent findings on the novel aspects of both synthases established in gene knockout and overexpression experiments, current preclinical pharmacology, and potential ways of gene expression regulation. Identification of protein-protein interactions and post-translational modifications appear to be the main options for modulating PTGIS and TBXAS1 activity. The microsomal CYPs are known to form complexes with each other and direct interactions of CYP2E1 with both synthases can probably lead to modulation of their activity. Progress in the preclinical development of low molecular weight compounds as inhibitors of TBXAS1 is more prospective than PTGIS that is applied as gene therapy biologicals for <em>in vivo</em> production of prostacyclin due to its noticeable anticancer and vasodilator effects.</div></div>","PeriodicalId":251,"journal":{"name":"Biochimie","volume":"234 ","pages":"Pages 76-88"},"PeriodicalIF":3.3,"publicationDate":"2025-04-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143850467","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Phenotypic changes associated with continuous long term in vitro expansion of bone marrow-derived mesenchymal stem cells 与骨髓间充质干细胞体外连续长期扩增相关的表型变化

IF 3.3 3区生物学

Biochimie Pub Date : 2025-04-08 DOI: 10.1016/j.biochi.2025.04.002

Vitali V. Maldonado , Hanna Jensen , C. Lowry Barnes , Rebekah M. Samsonraj

{"title":"Phenotypic changes associated with continuous long term in vitro expansion of bone marrow-derived mesenchymal stem cells","authors":"Vitali V. Maldonado , Hanna Jensen , C. Lowry Barnes , Rebekah M. Samsonraj","doi":"10.1016/j.biochi.2025.04.002","DOIUrl":"10.1016/j.biochi.2025.04.002","url":null,"abstract":"<div><div>In vitro expansion of mesenchymal stem cells is necessary to obtain a higher cell number for clinical applications. However, long-term expansion can produce significant phenotypic changes on these cells, decreasing their therapeutic utility. Therefore, understanding the phenotypic changes that long-term expansion triggers in mesenchymal stem cells will allow for better and more consistent cell therapy results. Here, we evaluate the phenotypic changes caused by continuous passaging through colony forming unit-fibroblast assay, senescence beta-galactosidase staining, morphology examination, secretome analysis, surface marker expression, protein quantification, osteogenic and adipogenic differentiation, and CD4<sup>+</sup> T lymphocyte immunosuppressive potential. Long-term in vitro culture decreases mesenchymal stem cell osteogenic potential and self-renewal, increases cell size, and senescence, but does not consistently affect adipogenic differentiation. Surface marker expression remains similar for positive and negative markers, while secretory phenotype shifts with decreased p14ARF, MMP-3, p21 Waf1/Cip1,ENA-78, GCP-2, GROα, IL-3, IL-7, IL-8, RANTES, TNFβ, and VEGF-A expression, and increased p53, p16 INK4a, MCP-1, and SDF-1 expression. Immunomodulatory potential remains unchanged. These findings can help better understand the phenotypic changes that mesenchymal stem cells undergo while expanded in vitro.</div></div>","PeriodicalId":251,"journal":{"name":"Biochimie","volume":"234 ","pages":"Pages 62-75"},"PeriodicalIF":3.3,"publicationDate":"2025-04-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143848046","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Divergent ferroptotic pathways in breast cancer cells: IGFBP6-regulated mitochondrial lipid peroxidation under erastin and omega-3 DHA treatment 乳腺癌细胞中不同的铁细胞凋亡途径：igfbp6在erastin和omega-3 DHA处理下调节线粒体脂质过氧化。

IF 3.3 3区生物学

Biochimie Pub Date : 2025-04-01 DOI: 10.1016/j.biochi.2025.03.010

Mariia Silkina , Alexandra Razumovskaya , Timur Kulagin , Artem Fatkulin , Karina Klycheva , Darya Olkhovik , Darya Averinskaya , Oksana Kolodeeva , Olga Kolodeeva , Alexander Tonevitsky , Sergey Nikulin

{"title":"Divergent ferroptotic pathways in breast cancer cells: IGFBP6-regulated mitochondrial lipid peroxidation under erastin and omega-3 DHA treatment","authors":"Mariia Silkina , Alexandra Razumovskaya , Timur Kulagin , Artem Fatkulin , Karina Klycheva , Darya Olkhovik , Darya Averinskaya , Oksana Kolodeeva , Olga Kolodeeva , Alexander Tonevitsky , Sergey Nikulin","doi":"10.1016/j.biochi.2025.03.010","DOIUrl":"10.1016/j.biochi.2025.03.010","url":null,"abstract":"<div><div>Breast cancer remains a major challenge and new therapeutic approaches are needed for its treatment. Ferroptosis is considered a promising alternative cell death mechanism to eliminate resistant cancer cells. In previous works, we identified that lower <em>IGFBP6</em> gene expression in tumor tissue corresponds to a worse prognosis for breast cancer patients and, at the same, time makes them more sensitive to ferroptosis. In this study, we further investigated the mechanism of ferroptosis induction in <em>IGFBP6</em> knockdown and control MDA-MB-231 breast cancer cells by the canonical ferroptosis inducer erastin and omega-3 docosahexaenoic acid (DHA). Our results indicate that there is a significant overlap between the mechanisms of action of both of these molecules, as they regulate the same subset of genes, and their action can be inhibited by canonical ferroptosis inhibitors. On the other hand, we also observed significant differences between the effects of erastin and DHA. The most notable of these are the additional activation of apoptosis-related genes by DHA and its minor peroxidation of mitochondrial lipid membranes. Interestingly, our kinetic analysis of ferroptosis induction showed that <em>IGFBP6</em> knockdown cells began to die earlier and could hardly be rescued from erastin-induced ferroptosis by mitochondrial antioxidant SkQ1, in contrast to control cells. Overall, our data suggest that the action of DHA is less dependent on mitochondrial membrane peroxidation during ferroptosis induction, and this molecule can be a promising candidate for the treatment of breast cancer, especially in the case of reduced <em>IGFBP6</em> gene expression in cancer cells.</div></div>","PeriodicalId":251,"journal":{"name":"Biochimie","volume":"234 ","pages":"Pages 48-61"},"PeriodicalIF":3.3,"publicationDate":"2025-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143782270","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Molecular and biochemical approaches of the trypanothione system in Leishmania spp.: A key player in parasite resistance to antimonial therapy 利什曼原虫中锥虫硫酮系统的分子和生化研究：对抗疟治疗产生抗性的关键因素。

IF 3.3 3区生物学

Biochimie Pub Date : 2025-03-25 DOI: 10.1016/j.biochi.2025.03.007

Geovane Dias-Lopes , Sara Maria Xavier Cruz , Bernardo Acácio Santini Pereira , Anabel Zabala-Peñafiel , Carlos Roberto Alves

{"title":"Molecular and biochemical approaches of the trypanothione system in Leishmania spp.: A key player in parasite resistance to antimonial therapy","authors":"Geovane Dias-Lopes , Sara Maria Xavier Cruz , Bernardo Acácio Santini Pereira , Anabel Zabala-Peñafiel , Carlos Roberto Alves","doi":"10.1016/j.biochi.2025.03.007","DOIUrl":"10.1016/j.biochi.2025.03.007","url":null,"abstract":"<div><div>The trypanothione system is a crucial antioxidant defense mechanism in <em>Leishmania</em> spp. The enzymes involved, including trypanothione reductase (TR), trypanothione synthetase (TS), tryparedoxin (TXN) and tryparedoxin peroxidase (TXNPx) are essential for maintaining the redox balance. This system plays a fundamental role in the biology of <em>Leishmania</em> spp., contributing to parasite resistance against metalloid-based treatments, such as trivalent antimony (Sb<sup>3+</sup>). The mechanisms underlying this resistance, particularly those linked to the functionality of the trypanothione system, have garnered increasing interest. This review prioritizes studies conducted with clinical isolates of <em>Leishmania</em> spp. that evaluated gene expression, protein abundance, and enzyme activity to determine how variations in trypanothione-related mechanisms influence their clinical outcomes. Additionally, complementary strategy involving different protocols to determine intracellular non-protein thiols have further enrich the information into these studies. Notably, the evidence gathered here highlights that studies have focused on only four <em>Leishmania</em> spp. with just one belonging to the <em>Viannia</em> subgenus. Several approaches have been used to determine TR and TXNPx enzyme activity in parasite lysates, supporting their use as tools for studying resistant phenotypes. Additionally, the assessment of TR, TXNPx and TS activities has been applied in kinetic studies for screening of inhibitor compounds. The functional insights presented herein may aid in elucidating the basis of parasite resistance and guide the development of more effective therapeutic strategies against leishmaniasis in its different clinical forms.</div></div>","PeriodicalId":251,"journal":{"name":"Biochimie","volume":"234 ","pages":"Pages 40-47"},"PeriodicalIF":3.3,"publicationDate":"2025-03-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143733614","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Glutathionylation and metabolic dysfunction-associated steatotic liver disease 谷胱甘肽化与代谢功能障碍相关的脂肪变性肝病。

IF 3.3 3区生物学

Biochimie Pub Date : 2025-03-25 DOI: 10.1016/j.biochi.2025.03.006

Zhe Jiang , Lin Chen , Xiaobing Dou

引用次数: 0

Leishmania (Viannia) braziliensis Thor strain and subpopulations Thor03, Thor10, and Thor22 have differences in the surface membrane proteases activity profile 巴西利什曼原虫（Viannia） Thor菌株及其亚群Thor 03、Thor10和Thor 22在表面膜蛋白酶活性谱上存在差异。

IF 3.3 3区生物学

Biochimie Pub Date : 2025-03-23 DOI: 10.1016/j.biochi.2025.03.005

Fatemeh Farshchi , Geovane Dias-Lopes , Luzia Monteiro de Castro Cortes , Léa Cysne-Finkelstein , Franklin Souza-Silva , Carlos Roberto Alves

{"title":"Leishmania (Viannia) braziliensis Thor strain and subpopulations Thor03, Thor10, and Thor22 have differences in the surface membrane proteases activity profile","authors":"Fatemeh Farshchi , Geovane Dias-Lopes , Luzia Monteiro de Castro Cortes , Léa Cysne-Finkelstein , Franklin Souza-Silva , Carlos Roberto Alves","doi":"10.1016/j.biochi.2025.03.005","DOIUrl":"10.1016/j.biochi.2025.03.005","url":null,"abstract":"<div><div>The <em>Leishmania (Viannia) braziliensis</em> Thor strain is composed of subpopulations with distinct biological features, as differences of the virulence profile <em>in vitro</em> and <em>in vivo</em> in murine model. As the surface of these parasites is the first contact with the host, this study assesses comparative approaches of surface membrane proteases of promastigotes and axenic amastigotes of <em>L. (V.) braziliensis</em> Thor strain and Thor03, Thor10, and Thor22 subpopulations, accessing differential profiles among these parasites. Here is explored the phospholipase C (PLC) property as a pivotal tool to selectively recover surface proteases of these parasites. The treatment of parasites with PLC yielded protein fractions with metalloprotease, cysteine protease, and serine protease activities, which were detectable by gelatin-SDS-PAGE and fluorogenic substrates and specific inhibitors, showing distinct profiles from both promastigotes and axenic amastigotes of the Thor strain, Thor03, Thor10, and Thor22 subpopulations. Data of protease activity quantitative in solution show metalloprotease as the highest activity, followed by cysteine protease and serine protease onto the surface of promastigotes and axenic amastigotes. The biological significance of these findings points to the potential of the Thor strain, helped by respective subpopulations, to adapt to hosts, as well as reinforcing the importance of this class of enzyme in the first hours of infection.</div></div>","PeriodicalId":251,"journal":{"name":"Biochimie","volume":"234 ","pages":""},"PeriodicalIF":3.3,"publicationDate":"2025-03-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143712474","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0