Biochimie最新文献_第4页

Both single mutation and molecular conjugation on the mutant prevent calcitonin from forming amyloid fibrils 突变体的单突变和分子偶联均可阻止降钙素形成淀粉样原纤维。

IF 3 3区生物学

Biochimie Pub Date : 2026-03-01 Epub Date: 2025-11-29 DOI: 10.1016/j.biochi.2025.11.012

Chia-Chi Chang , Govindan Sivakumar , Meng-Chieh Huang , Yun-Hsuan Chen , Shan-Rong Wu , Yun-Ju Lai , Chian-Hui Lai , Ling-Hsien Tu

{"title":"Both single mutation and molecular conjugation on the mutant prevent calcitonin from forming amyloid fibrils","authors":"Chia-Chi Chang , Govindan Sivakumar , Meng-Chieh Huang , Yun-Hsuan Chen , Shan-Rong Wu , Yun-Ju Lai , Chian-Hui Lai , Ling-Hsien Tu","doi":"10.1016/j.biochi.2025.11.012","DOIUrl":"10.1016/j.biochi.2025.11.012","url":null,"abstract":"<div><div>Human calcitonin (hCT) is a peptide hormone made up of 32 amino acids, produced by the parafollicular cells of the thyroid gland. Its main role is to decrease blood calcium levels by inhibiting osteoclast activity and reducing calcium reabsorption in the kidneys and intestines. This characteristic positions hCT as a potential treatment for bone-related conditions, such as osteoporosis and Paget's disease. However, hCT has a strong tendency to form amyloid fibrils, which can lead to a loss of its biological function. This research aims to stabilize hCT's conformation through molecular conjugation to prevent its aggregation. We used a cross-linking tool with a boronic ester structure (<strong>NPC</strong>) to react with the side-chain amino groups of two residues in hCT, effectively \"locking\" its structure. hCT can be \"unlocked\" using hydrogen peroxide, returning it to its original form. To support this experimental design, we synthesized two hCT variants, hCT-Q14K and hCT-F22K. Our results showed that both variants successfully conjugated with <strong>NPC</strong>, and the conjugation could be easily removed with hydrogen peroxide within 30 min. <strong>NPC</strong>-conjugated hCT did not form amyloid as revealed by transmission electron microscopy. Moreover, thioflavin-T fluorescence kinetic studies and dynamic light scattering measurement demonstrated that the aggregation tendencies of hCT-F22K were significantly lower than those of hCT, while still maintaining their biological activity, suggesting this design is feasible. This approach may significantly aid in the development of calcitonin drugs.</div></div>","PeriodicalId":251,"journal":{"name":"Biochimie","volume":"242 ","pages":"Pages 1-8"},"PeriodicalIF":3.0,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145650371","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

Giardia duodenalis: Base excision repair pathway enzyme FEN1 carries out catalytic activities pertaining to NER pathway 十二指肠贾第虫：碱基切除修复途径酶FEN1具有与NER途径相关的催化活性。

IF 3 3区生物学

Biochimie Pub Date : 2026-03-01 Epub Date: 2025-12-24 DOI: 10.1016/j.biochi.2025.12.010

Ulises Omar García-Lepe, Sofía Gabriela Tomás-Morales, María Teresa Izaguirre-Hernández, María Luisa Bazán-Tejeda, Rosa María Bermúdez-Cruz

{"title":"Giardia duodenalis: Base excision repair pathway enzyme FEN1 carries out catalytic activities pertaining to NER pathway","authors":"Ulises Omar García-Lepe, Sofía Gabriela Tomás-Morales, María Teresa Izaguirre-Hernández, María Luisa Bazán-Tejeda, Rosa María Bermúdez-Cruz","doi":"10.1016/j.biochi.2025.12.010","DOIUrl":"10.1016/j.biochi.2025.12.010","url":null,"abstract":"<div><div><em>Giardia duodenalis</em> is a binuclear protozoan that causes intestinal infection in humans and animals. The life cycle of <em>G. duodenalis</em> is comprised by 2 stages: trophozoite (vegetative, ploidy: 4 N) and cyst (infective, ploidy: 8–16 N) and the transition from one to another requires a precise coordination as well as the support of the DNA repair machinery. While DNA homologous recombination DNA repair has recently been characterized, NER and BER are pathways that had not been explored. Most of the structure specific enzymes (SSE) participate in a variety of processes like DNA replication stress, DNA adduct repair and Holliday junction processing. In an effort to explore these kinds of enzymes in <em>G.</em> duodenalis, a minimalist parasite, we aimed at characterizing the Fen1 enzyme by cloning its gene to study its catalytic properties (binding and nuclease) using flap and bubble DNA substrates. Unexpectedly, we found that GdFen-1 is able to cleave bubble DNA, then to shed light on which domains of this enzyme are responsible for this activity, giardial acid block and a portion of a cap region were substituted by their human counterparts, and while acid block substitution did not affect this activity, the modification in the cap region did. The possible implications of these findings are addressed.</div></div>","PeriodicalId":251,"journal":{"name":"Biochimie","volume":"242 ","pages":"Pages 85-96"},"PeriodicalIF":3.0,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145844659","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

Mechanistic insights into NOBA hydrolysis by viper venom secreted phospholipase A2 毒蛇毒液分泌的磷脂酶A2水解NOBA的机理

IF 3 3区生物学

Biochimie Pub Date : 2026-03-01 Epub Date: 2025-12-05 DOI: 10.1016/j.biochi.2025.12.003

Ana Rita Calixto , Roberto Pinto , Maciej Spiegel , Maria João Ramos , Pedro Alexandrino Fernandes

{"title":"Mechanistic insights into NOBA hydrolysis by viper venom secreted phospholipase A2","authors":"Ana Rita Calixto , Roberto Pinto , Maciej Spiegel , Maria João Ramos , Pedro Alexandrino Fernandes","doi":"10.1016/j.biochi.2025.12.003","DOIUrl":"10.1016/j.biochi.2025.12.003","url":null,"abstract":"<div><div>Snake envenoming remains a major global health challenge, particularly in tropical and subtropical regions. Among the key toxic components of snake venom, secreted phospholipases A<sub>2</sub> (sPLA<sub>2</sub>) play a crucial role by hydrolysing cell membrane phospholipids, leading to membrane disruption and severe toxic effects such as inflammation, neurotoxicity, and myotoxicity.</div><div>To study sPLA<sub>2</sub> catalytic activity, synthetic soluble substrates like 4-nitro-3-octanoyloxy benzoic acid (NOBA) are widely used in experimental assays, in alternative to membrane phospholipids. However, it is questionable whether mechanistic conclusions taken with small, soluble substrates can be extrapolated to true cell-membrane substrates.</div><div>Here, we employed QM/MM calculations to investigate the catalytic mechanism of sPLA<sub>2</sub> using NOBA as a substrate. Our focus was on a sPLA<sub>2</sub> from <em>Bothrops asper</em>. However, the high conservation of sPLA<sub>2</sub> active sites suggests our conclusions should be generalisable to the sPLA<sub>2</sub> of most snake species.</div><div>The results reveal a mixed single-water/assisted water mechanism. First, a water molecule, deprotonated by His<sub>47</sub>, performs a nucleophilic attack on NOBA's carbonyl carbon, with a free energy barrier of 12.8 kcal/mol, resembling the single-water pathway. The collapse of the tetrahedral intermediate and protonation of the leaving group involves two water molecules, resembling the assisted-water pathway. This mixed pathway highlights the catalytic versatility of sPLA<sub>2</sub> and offers new insights into its enzymatic activity with synthetic substrates. Importantly, our finding support NOBA as a valid and suitable substrate for studying the chemical component of sPLA<sub>2</sub> toxicity, even though it does not account for the equally important membrane-binding component of the toxicity mechanism.</div></div>","PeriodicalId":251,"journal":{"name":"Biochimie","volume":"242 ","pages":"Pages 108-117"},"PeriodicalIF":3.0,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145703245","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

Dietary modulation of purine metabolism and uric acid homeostasis in cancer patients with an ileostomy 回肠造口术后癌症患者嘌呤代谢和尿酸稳态的饮食调节。

IF 3 3区生物学

Biochimie Pub Date : 2026-03-01 Epub Date: 2026-01-05 DOI: 10.1016/j.biochi.2026.01.002

Kamil Aleksander Sobieszek , Jakub Frankowski , Mateusz Labudda

{"title":"Dietary modulation of purine metabolism and uric acid homeostasis in cancer patients with an ileostomy","authors":"Kamil Aleksander Sobieszek , Jakub Frankowski , Mateusz Labudda","doi":"10.1016/j.biochi.2026.01.002","DOIUrl":"10.1016/j.biochi.2026.01.002","url":null,"abstract":"<div><div>Nutritional management in cancer patients with an ileostomy poses specific challenges due to impaired nutrient absorption, elevated metabolic demands, and the need to control serum urate levels. This review examines the biochemical and clinical relevance of plant-based foods in addressing these issues, with particular emphasis on purine content, digestibility, and metabolic outcomes. Current evidence shows that legumes, soy products, nuts, seeds, whole grains, vegetables, fruits, and fortified foods generally contain low to moderate purine levels and are well tolerated by ileostomy patients. Incorporating these foods into individualized dietary plans supports adequate protein and micronutrient intake, enhances tissue repair, and reduces the risk of gout flares without compromising gastrointestinal function. Moreover, plant-derived bioactive compounds and antioxidants may mitigate inflammation and oxidative stress associated with cancer progression and hyperuricemia. Collectively, a carefully designed plant-based diet can meet the nutritional needs of cancer patients with an ileostomy while contributing to effective gout management and improved metabolic homeostasis.</div></div>","PeriodicalId":251,"journal":{"name":"Biochimie","volume":"242 ","pages":"Pages 134-142"},"PeriodicalIF":3.0,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145919187","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

Inhibiting catalytic activity of Plasmodium falciparum aspartate protease plasmepsin V: A biochemical approach to malaria intervention 抑制恶性疟原虫天冬氨酸蛋白酶Plasmepsin V的催化活性：一种生物化学方法干预疟疾。

IF 3 3区生物学

Biochimie Pub Date : 2026-03-01 Epub Date: 2025-12-13 DOI: 10.1016/j.biochi.2025.12.006

Anitadevi K. Prajapati , Yogesh Kumar , Sinjan Choudhary

{"title":"Inhibiting catalytic activity of Plasmodium falciparum aspartate protease plasmepsin V: A biochemical approach to malaria intervention","authors":"Anitadevi K. Prajapati , Yogesh Kumar , Sinjan Choudhary","doi":"10.1016/j.biochi.2025.12.006","DOIUrl":"10.1016/j.biochi.2025.12.006","url":null,"abstract":"<div><div>Plasmepsin V, an aspartate protease, is crucial for the survival of the malarial parasite <em>Plasmodium falciparum</em>, making it a promising target for antimalarial drug development. In this study, the quinoline-based drugs amodiaquine (AQ) and clioquinol (CQ) are repurposed to inhibit the catalytic activity of <em>Pf</em>PlmV. Fluorescence quenching assays suggest that both AQ and CQ exhibit moderate binding affinity for <em>Pf</em>PlmV. Temperature-dependent thermodynamic analyses indicate the involvement of both H-bonding/electrostatic interactions and hydrophobic interactions in the binding of AQ/CQ with <em>Pf</em>PlmV. Isothermal titration calorimetry (ITC) indicates that these interactions are both enthalpically and entropically favourable. Molecular docking studies show that AQ binds directly to the substrate-binding site of <em>Pf</em>PlmV, engaging catalytic dyad residues Asp118 and Asp365, whereas CQ binds to <em>Pf</em>PlmV but does not directly interact with the catalytic dyad. Molecular dynamics simulations also corroborated the docking results, revealing that AQ forms a more conformationally stable and hydrophobic interactions driven complex with PfPlmV compared to CQ. Further, the enzyme kinetics studies using fluorogenic substrate demonstrated that AQ efficiently inhibits the catalytic activity of <em>Pf</em>PlmV with an IC<sub>50</sub> value of (0.42 ± 0.04) μM via competitive inhibition mode. In contrast, CQ fails to bind at the catalytic site and does not exhibit any inhibitory effect on <em>Pf</em>PlmV activity. These mechanistic insights will lay the foundations for new biochemical approaches to develop targeted therapies aimed at malaria disease.</div></div>","PeriodicalId":251,"journal":{"name":"Biochimie","volume":"242 ","pages":"Pages 42-58"},"PeriodicalIF":3.0,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145758705","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

Sodium benzoate, a common food preservative-induced Insulin resistance and atrophy in C2C12 myotubes 苯甲酸钠是一种常见的食品防腐剂，可诱导胰岛素抵抗和C2C12肌管萎缩。

IF 3 3区生物学

Biochimie Pub Date : 2026-02-01 Epub Date: 2025-11-18 DOI: 10.1016/j.biochi.2025.11.009

Onkar Sharma , Anita Dua , Sanjeev Gupta , Elisha Injeti , Ashwani Mittal

{"title":"Sodium benzoate, a common food preservative-induced Insulin resistance and atrophy in C2C12 myotubes","authors":"Onkar Sharma , Anita Dua , Sanjeev Gupta , Elisha Injeti , Ashwani Mittal","doi":"10.1016/j.biochi.2025.11.009","DOIUrl":"10.1016/j.biochi.2025.11.009","url":null,"abstract":"<div><div>Sodium benzoate (SB), a widely used food preservative, has been reported to possess several therapeutic benefits including treating neurodegenerative disorders as well as negative effects such as altering glucose homeostasis. However, limited attention has been given to its role in inducing insulin resistance (IR). Therefore, current study was designed to investigate the relationship between SB and insulin resistance using C2C12 myotubes and to explore the possible underlying mechanisms. Myotubes were exposed to SB (20 mM) for 24 h and glucose consumption and uptake assays along with confocal microscopy (GLUT4 translocation) were used to evaluate IR. Oxidative stress indicators <em>i.e.</em> reactive oxygen species (ROS), lipid peroxidation (LPO), and reduced glutathione (GSH) levels were also measured. Myotubes morphology along with atrophic markers (calpain activity) were evaluated. Data show that in C2C12 myotubes, SB treatment reduced glucose consumption and uptake in a dose-dependent manner. SB inducing IR by disrupting insulin-mediated GLUT4 translocation in cultured myotubes. Alteration in oxidative stress-related markers (<em>i.e.</em> elevated ROS and LPO levels, reduced GSH) were also observed in SB-treated myotubes. Furthermore, decrease in the fusion index, length, and diameter of myotubes along with upregulation of calpain activity and decrease in muscle protein were also observed. Study concludes that SB exposure not only induced IR but also caused atrophy and oxidative stress in the C2C12 myotubes.</div></div>","PeriodicalId":251,"journal":{"name":"Biochimie","volume":"241 ","pages":"Pages 56-71"},"PeriodicalIF":3.0,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145566653","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

Design of competitive inhibitory peptides for 3-hydroxy-3-Methylglutaryl coenzyme A reductase 3-羟基-3-甲基戊二酰辅酶A还原酶竞争性抑制肽的设计

IF 3 3区生物学

Biochimie Pub Date : 2026-02-01 Epub Date: 2025-11-01 DOI: 10.1016/j.biochi.2025.10.017

Valeriy V. Pak , Shomansur Sh Sagdullaev , Aleksandr V. Pak

{"title":"Design of competitive inhibitory peptides for 3-hydroxy-3-Methylglutaryl coenzyme A reductase","authors":"Valeriy V. Pak , Shomansur Sh Sagdullaev , Aleksandr V. Pak","doi":"10.1016/j.biochi.2025.10.017","DOIUrl":"10.1016/j.biochi.2025.10.017","url":null,"abstract":"<div><div>The effectiveness of statins in preventing hypercholesterolemia and associated cardiovascular diseases has been confirmed for a long time. Statins are competitive inhibitors of 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMGR). This enzyme plays a key role in the reaction that represents the rate-limiting step in cholesterol biosynthesis. Numerous studies on the properties of food-derived peptides have revealed various bioactivities that influence human health, including the modulation of endogenous cholesterol levels. Ongoing studies have shown that some peptides function using the same mechanism as statins and can be considered another class of compounds for HMGR inhibition.To date, the competitive inhibition of HMGR by peptides has been confirmed for 36 of them. The half-maximal inhibitory concentration (IC<sub>50</sub>) of the most active food-derived peptide was found to be 12.8 μM, which is significantly lower than statin activity. This review presents approaches for modeling peptides to enhance their activity. Analysis of the peptides' physicochemical characteristics revealed the potential to design a more active peptide by adjusting a single parameter. The most active designed peptide exhibited 700 times the activity of an isolated peptide found in food. These studies demonstrate the potential of using peptides to develop nutraceuticals or drugs that prevent hypercholesterolemia.</div></div>","PeriodicalId":251,"journal":{"name":"Biochimie","volume":"241 ","pages":"Pages 1-10"},"PeriodicalIF":3.0,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145440172","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

A Trichomonas vaginalis C2-XYPPX-repeat protein with a structured C2 domain displaying dampened flexibility upon binding calcium 阴道毛滴虫C2- xyppx -重复蛋白，具有结构化的C2结构域，在结合钙时显示受湿的灵活性。

IF 3 3区生物学

Biochimie Pub Date : 2026-02-01 Epub Date: 2025-11-22 DOI: 10.1016/j.biochi.2025.11.011

Garry W. Buchko , Lijun Liu , Kevin P. Battaile , Justin K. Craig , Elizabeth K. Harmon , Wesley C. Van Voorhis , Peter J. Myler , Scott Lovell

{"title":"A Trichomonas vaginalis C2-XYPPX-repeat protein with a structured C2 domain displaying dampened flexibility upon binding calcium","authors":"Garry W. Buchko , Lijun Liu , Kevin P. Battaile , Justin K. Craig , Elizabeth K. Harmon , Wesley C. Van Voorhis , Peter J. Myler , Scott Lovell","doi":"10.1016/j.biochi.2025.11.011","DOIUrl":"10.1016/j.biochi.2025.11.011","url":null,"abstract":"<div><div>C2 domains are ubiquitous membrane-binding modules of ∼130 residues in eukaryotes that are often associated with proteins involved in membrane trafficking and lipid modification. The genome of <em>Trichomonas vaginalis,</em> the most common, non-viral, sexually transmitted human pathogen, encodes eight genes that contain a N-terminal C2 module linked to a XYPPX-repeat domain of more than four XYPPX repeats (C2-XYPPX). While the function of the XYPPX-repeat domain remains unknown, its multiple association with C2 domains in <em>T. vaginalis</em> suggests it is important. The C2 domain from one of these C2-XYPPX-repeat proteins, <em>Tv</em>-C2-1, was structurally and physically characterized using X-ray crystallography and NMR spectroscopy. The crystal structure for <em>Tv</em>-C2-1 shows that this domain shares a fold common to all C2 domains, a compact Greek-key motif composed of eight anti-parallel β-strands in the type-2 topology. An NMR chemical shift perturbation study with Ca<sup>2+</sup> showed that <em>Tv</em>-C2-1 bound two Ca<sup>2+</sup> atoms primarily via two loops (loop-1 and loop-3) on the predicted calcium binding face of the protein with K<sub>d</sub>s of 58.0 ± 0.1 μM and 232 ± 6 μM. Estimations of the overall rotational correlation time, τ<sub>c</sub>, in the apo (11.1 ns) and Ca<sup>2+</sup>-bound (9.2 ns) state suggests the protein becomes more compact upon Ca<sup>2+</sup> binding, consistent with a decrease in dynamics in loop-3 and marginally in loop-1 suggested by amide <sup>15</sup>N heteronuclear steady-state {<sup>1</sup>H}-<sup>15</sup>N NOEs. Showing <em>Tv</em>-C2-1 binds calcium and adopts a compact Greek-key motif structure, two primary features of C2 domains, suggests understanding the function of the XYPPX-repeat domain may be warranted.</div></div>","PeriodicalId":251,"journal":{"name":"Biochimie","volume":"241 ","pages":"Pages 44-55"},"PeriodicalIF":3.0,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145597251","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

Paracrine effect of fibroblasts on the proliferation and differentiation of bovine satellite cells in vitro 成纤维细胞对体外牛卫星细胞增殖分化的旁分泌作用。

IF 3 3区生物学

Biochimie Pub Date : 2026-02-01 Epub Date: 2025-11-15 DOI: 10.1016/j.biochi.2025.11.006

Karolina Zygmunt , Kacper Żukowski , Katarzyna Piórkowska , Grzegorz Smołucha , Alicja Wierzbicka , Wojciech Witarski

{"title":"Paracrine effect of fibroblasts on the proliferation and differentiation of bovine satellite cells in vitro","authors":"Karolina Zygmunt , Kacper Żukowski , Katarzyna Piórkowska , Grzegorz Smołucha , Alicja Wierzbicka , Wojciech Witarski","doi":"10.1016/j.biochi.2025.11.006","DOIUrl":"10.1016/j.biochi.2025.11.006","url":null,"abstract":"<div><div>The co-culture of fibroblasts and muscle cells is essential for obtaining cultured meat using <em>in vitro</em> techniques. While fibroblasts are known to promote myogenesis through cell-cell interactions, their paracrine effects and associated transcriptomic changes remain unknown. This study aimed to investigate the paracrine effect of fibroblasts on myogenesis, focusing on transcriptome profiling. Muscle satellite cells (isolated enzymatically) and fibroblasts (isolated through the explant culture) were co-cultured in 0.4 μm transwell plates for 5 days of proliferation and 24 and 72 h of differentiation. RNA-Seq and protein analysis (Western Blot, ELISA, immunofluorescence) were used to assess changes in myogenic marker expression. RNA-Seq revealed changes in many genes involved in myogenesis, such as upregulation of EGR1, IL6, and SOCS3 and downregulation of ITGA7. ELISA showed significantly higher MyHC levels at the proliferation stage in the co-culture group (p = 0.0183), with no significant differences at differentiation stages. To summarize, fibroblasts promote early myogenic differentiation and could modulate the extent of myogenic differentiation.</div></div>","PeriodicalId":251,"journal":{"name":"Biochimie","volume":"241 ","pages":"Pages 26-43"},"PeriodicalIF":3.0,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145543953","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

Are Traditional mutant controls sufficient to identify true RNA G-quadruplex binding proteins? 传统的突变控制是否足以识别真正的RNA g -四重结合蛋白？

IF 3 3区生物学

Biochimie Pub Date : 2026-02-01 Epub Date: 2025-11-01 DOI: 10.1016/j.biochi.2025.10.018

Marc-Antoine Turcotte, Louise Dao Josépha Crespo, Jean-Pierre Perreault

{"title":"Are Traditional mutant controls sufficient to identify true RNA G-quadruplex binding proteins?","authors":"Marc-Antoine Turcotte, Louise Dao Josépha Crespo, Jean-Pierre Perreault","doi":"10.1016/j.biochi.2025.10.018","DOIUrl":"10.1016/j.biochi.2025.10.018","url":null,"abstract":"<div><div>G-quadruplexes (G4s) are stable non-canonical RNA structures involved in various regulatory processes, whose recognition by G4-binding proteins (G4BPs) is often studied using affinity purification and biochemical validation. A critical factor in validating true G4BPs lies in the choice of negative controls, which are frequently limited to simple G-to-A point mutations. Here, we reassess the classification of Guanine Nucleotide-Binding Protein-Like 1 (GNL1), previously identified as an RNA G4BP (rG4BP), by employing 7-deazaguanine (7dG) substitutions and salt variation strategies. Using fluorescence assays with N-methyl mesoporphyrin IX (NMM) and electrophoretic mobility shift assays (EMSAs), we show that GNL1 binds wild-type and 7dG-modified RNAs with comparable affinities. Additional experiments using potassium and lithium ions further that GNL1 binding is independent of G4 structure. Finally, truncation of PRKN RNA to its G4 core significantly reduced GNL1 binding, indicating that the protein does not interact with the folded G4 itself. These results collectively demonstrate that GNL1 binds guanine-containing sequences rather than true G4s, and that its prior classification as a G4BP was likely due to insufficient controls. Our findings highlight the importance of using robust negative controls, such as 7dG substitution and ionic condition modulation, in the accurate identification of <em>bona fide</em> G4BPs.</div></div>","PeriodicalId":251,"journal":{"name":"Biochimie","volume":"241 ","pages":"Pages 11-15"},"PeriodicalIF":3.0,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145440181","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