Analytical biochemistry最新文献

{"title":"Flower-like biomimetic enzyme for rapid and sensitive detection of zearalenone in vegetable oil deodorizer distillate","authors":"Lin Zhiyuan ,&nbsp;Xue Minqiao ,&nbsp;Xia Jiaojiao ,&nbsp;Suo Hongbo ,&nbsp;Huang Rubing ,&nbsp;Zou Bin","doi":"10.1016/j.ab.2025.115780","DOIUrl":"10.1016/j.ab.2025.115780","url":null,"abstract":"<div><div>In order to achieve high quality production of vitamin E and plant sterols, it is necessary to conduct rapid and accurate detection of fungal toxins in their production raw material (vegetable oil deodorizer distillate, VODD). In this study, the flower-like biomimetic enzyme of silver-doped ZnO was synthesized through wet chemical method and in-situ reduction method. Based on above work, a flower-like biomimetic enzyme modified glass carbon electrode was fabricated, and its excellent detection capability against fungal toxins zearalenone was confirmed through electrochemical analysis. The detection limit was 8 ng mL⁻¹, with a linear range of 40 ng mL⁻¹-25 μg mL⁻¹. Simultaneously, the biomimetic enzyme sensor takes only 10 min from preparation to completion of detection, and the RSD between the 7 repeated test results was only 0.612 %. After seven days of storage, the current response value remains 91.5 % of the initial value. In practical applications, the recovery rate of zearalenone in VODD using this sensor ranged from 98.1 % to 102.08 %, yielding satisfactory results. Therefore, the novel flower-like biomimetic enzyme represents an ideal choice for developing zearalenone sensors and holds promising prospects for wide application in fungal toxins analysis.</div></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":"700 ","pages":"Article 115780"},"PeriodicalIF":2.6,"publicationDate":"2025-01-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143057793","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Different manipulations for resolving the overlapped spectra of novel anti-asthmatic combination: Green-blue-white triple evaluation tools","authors":"Michael Gamal Fawzy ,&nbsp;Hawa Khalil ,&nbsp;Alaa Reda ,&nbsp;Eman A. Bahgat","doi":"10.1016/j.ab.2025.115783","DOIUrl":"10.1016/j.ab.2025.115783","url":null,"abstract":"<div><div>This work represents different spectrophotometric techniques for concurrent quantification of Indacaterol (IND) and Mometasone furoate (MOM); co-formulated inhalation capsules to control asthma symptoms. Direct spectrophotometric (D⁰) approach was applied for IND assay. While, absorption factor (AF), ratio difference (RD), mean centering of the ratio spectra (MC), and continuous wavelet transform (CW) techniques were utilized for MOM quantification. The applied methods' validation was accomplished relative to the ICH guidelines. Within the linearity range of (4–20 μg/mL), these approaches provided accurate and precise assessment of IND and MOM in laboratory-synthetic mixtures and pharmaceutical formulation with determination coefficients higher than 0.9997. Moreover, the proposed methods demonstrated good detection and quantification levels. The respective LOD and LOQ for IND were (0.33 μg/mL) and (1.01 μg/mL). While, the LOD and LOQ for MOM were ranging from (0.16–0.30 μg/mL) and (0.47–0.91 μg/mL), respectively. Statistical comparison revealed no significant differences between the findings derived from the proposed methods and the reported one. The applied methods’ environmental effect was cheeked using Analytical Greenness Calculator (AGREE), Blue Applicability Grade Index (BAGI), and Red-Green-Blue 12 (RGB12). All the utilized metrics showed reliable scores, approving that the proposed methods were adhered to the sustainability principles.</div></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":"700 ","pages":"Article 115783"},"PeriodicalIF":2.6,"publicationDate":"2025-01-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143035973","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Identification of lipid metabolism-related gene markers and construction of a diagnostic model for multiple sclerosis: An integrated analysis by bioinformatics and machine learning","authors":"Fangjie Yang ,&nbsp;Xinmin Li ,&nbsp;Jing Wang ,&nbsp;Zhenfei Duan ,&nbsp;Chunlin Ren ,&nbsp;Pengxue Guo ,&nbsp;Yuting Kong ,&nbsp;Mengyao Bi ,&nbsp;Yasu Zhang","doi":"10.1016/j.ab.2025.115781","DOIUrl":"10.1016/j.ab.2025.115781","url":null,"abstract":"<div><h3>Background</h3><div>Multiple sclerosis (MS) is an autoimmune inflammatory disorder that causes neurological disability. Dysregulated lipid metabolism contributes to the pathogenesis of MS. This study aimed to identify lipid metabolism-related gene markers and construct a diagnostic model for MS.</div></div><div><h3>Methods</h3><div>Gene expression profiles for MS were obtained from the Gene Expression Omnibus database. Differentially expressed lipid metabolism-related genes (LMRGs) were identified and performed functional enrichment analysis. Least absolute shrinkage and selection operator (LASSO), random forest (RF), and protein-protein interaction (PPI) analysis were employed to screen hub genes. The predictive power of hub genes was evaluated using receiver operating characteristic (ROC) curves. We developed an artificial neural network (ANN) model and validated its performance in three test sets. Immune cell infiltration analysis, Gene set enrichment analysis, and ceRNA network construction were performed to explore the role of lipid metabolism in the pathogenesis of MS. Drugs prediction and molecular docking were utilized to identify potential therapeutic drugs.</div></div><div><h3>Results</h3><div>We identified 40 differentially expressed LMRGs, with significant enrichment in Arachidonic acid metabolism, Steroid hormone biosynthesis, Fatty acid elongation, and Sphingolipid metabolism. AKR1C3, NFKB1, and ABCA1 were identified as gene markers for MS, and their expression was upregulated in the MS group. The areas under the ROC curve (AUCs) for AKR1C3, NFKB1, and ABCA1 in the training set were 0.779, 0.703, and 0.726, respectively. The ANN model exhibited good discriminative ability in both the training and test sets, achieving an AUC of 0.826 on the training set and AUC values of 0.822, 0.890, and 0.833 on the test sets. Gamma.delta.T.cell, Natural.killer.T.cell, Plasmacytoid.dendritic.cell, Regulatory.T.cell, and Type.1.T.helper.cell were highly expressed in the MS group. A ceRNA network showed a complex regulatory interplay involving hub genes. Luteolin, isoflavone, and thalidomide had good binding affinities to the hub genes.</div></div><div><h3>Conclusion</h3><div>Our study emphasized the crucial role of lipid metabolism in MS, identifing AKR1C3, NFKB1, and ABCA1 as gene markers. The ANN model exhibited good performance on both the training and testing sets. These findings offer valuable insights into the molecular mechanisms underlying MS, and establish a scientific foundation for future research.</div></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":"700 ","pages":"Article 115781"},"PeriodicalIF":2.6,"publicationDate":"2025-01-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143035974","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"In-vivo implementation of the HPTLC methodology to rat plasma for the concurrent determination of a novel combination therapy for the management of COVID-19 (favipiravir and nitazoxanide)","authors":"Fadwa H. Edrees ,&nbsp;Maha M. Abdelrahman ,&nbsp;Amal B. Ahmed","doi":"10.1016/j.ab.2025.115779","DOIUrl":"10.1016/j.ab.2025.115779","url":null,"abstract":"<div><div>The 2019 coronavirus outbreak has prompted scientists to investigate pharmaceuticals to prevent the spread of the disease. Favipiravir (FAV) has received Food and Drug Administration FDA approval for the treatment of various viral infections with notable efficacy in clinical trials for COVID-19. Nitazoxanide (NTZ) is a broad-spectrum antiparasitic and antiviral agent used for the treatment of parasitic illnesses. Recently, the antiviral medications FAV and NTZ have been utilized therapeutically as early antiviral combination therapy for COVID-19, highlighting their safety, efficacy, and immunomodulatory effects. Despite several clinical studies on both FAV and NTZ, no analytical method has been developed for their concurrent detection. The objective of this study is to develop a TLC-densitometric method for their assessment and application to rat plasma. FAV, NTZ, and tizoxanide (the active metabolite of nitazoxanide (TZM)) were simultaneously determined using an HPTLC method embracing sulfasalazine as an internal standard (IS). Silica gel 60 F254 used as the stationary phase for chromatographic separation, the mobile phase consisted of chloroform-methanol-formic acid (9.5:0.5:0.2, v/v/v), with UV detection at 230 nm demonstrating linearity within the range of 0.5–5 μg/band. The proposed methodology has been shown to effectively measure the analyzed elements in both pure form and in-vivo rat plasma.</div></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":"700 ","pages":"Article 115779"},"PeriodicalIF":2.6,"publicationDate":"2025-01-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142998792","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Rapid, flexible fabrication of a microfluidic electrochemical chip nucleic acid target for selective, label-free detection of influenza virus DNA using catalytic redox-recycling","authors":"Hosna Ehzari ,&nbsp;Masoud Amiri ,&nbsp;Rahman Hallaj ,&nbsp;Marzieh Sadeghi","doi":"10.1016/j.ab.2025.115771","DOIUrl":"10.1016/j.ab.2025.115771","url":null,"abstract":"<div><div>H5N1 flu is a highly virulent and variable subtype of influenza with significant epidemic and pandemic potential. In this study, we introduce a novel, maskless, and rapid manufacturing process for a microfluidic chip integrated with electrodes for the quantitative detection of H5N1-DNA sequences. This detection leverages a catalytic redox-recycling signal via a novel Fe₃O₄@TMU-8 nanocomposite, which facilitates the turnover of the oxidation state of [Ru(NH₃)₆]³⁺, thereby amplifying the electrochemical signal output. The positively charged [Ru(NH₃)₆]³⁺ molecule associates with the phosphate backbone of the nucleic acids in H5N1-DNA. Changes in the aptasensor's redox-recycling signal, due to the hybridization of DNA sequences with [Ru(NH₃)₆]³⁺, were used as the electrochemical sensing response. Under optimal conditions, the signal exhibited a linear relationship with H5N1-DNA concentration, ranging from 1 fM to 1 nM, with a detection limit of 0.16 fM. This report details the fabrication of the microfluidic device using Poly(methyl methacrylate) (PMMA) sheet substrates. A laser system was employed to generate microfluidic patterns directly on the PMMA sheet. This biosensing device demonstrated long-term stability and good reproducibility, making it suitable for the quantitative assay of H5N1-DNA sequences. The results from food sample analyses further confirmed the applicability and effectiveness of the resulting biosensor.</div></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":"700 ","pages":"Article 115771"},"PeriodicalIF":2.6,"publicationDate":"2025-01-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142998793","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Innovative determination of Let-7a for lung cancer diagnosis using a duplex specific nuclease (DSN)-based electrochemical biosensor","authors":"Jianhao Xu,&nbsp;Haifeng Li","doi":"10.1016/j.ab.2025.115770","DOIUrl":"10.1016/j.ab.2025.115770","url":null,"abstract":"<div><div>In this study, we emphasize the importance of identifying Let-7a, a microRNA that is key in diagnosing and predicting lung cancer outcomes. Let-7a′s function as a biomarker is essential, as it affects tumor suppression and controls cell differentiation and growth. We developed a novel device, an electrochemical biosensor based on Duplex Specific Nuclease (DSN), that is designed for the accurate detection of Let-7a. This biosensor has a very low detection limit of 3.9 aM, showing its high sensitivity. The design is easy to use, requiring little training to operate. Its small size and portability enable the possibility of bedside and home use, which is a significant improvement for personalized healthcare. This versatility of the biosensor is very promising, as it can be applied to other disease biomarkers besides lung cancer. We expect this technology to improve disease diagnosis and patient recovery tracking, playing an important role in the future of medical diagnostics. The use of such sensitive and specific biosensors can transform the way diseases are managed, providing timely and precise information on disease progression and treatment effectiveness. The potential for this technology to help advance personalized medicine and patient care is huge, creating new opportunities in medical diagnostics and therapy monitoring.</div></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":"700 ","pages":"Article 115770"},"PeriodicalIF":2.6,"publicationDate":"2025-01-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142998791","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Microplate spectrophotometric method for regioselective lipase screening using structured triglycerides with punicic acid as probe","authors":"Enrique Ordaz ,&nbsp;Osvaldo Gómez-Secundino ,&nbsp;Hiram Y. Guerrero-Elias ,&nbsp;M. Angeles Camacho-Ruiz ,&nbsp;Ruben Espinosa-Salgado ,&nbsp;Antonio Escobedo-Reyes ,&nbsp;Juan C. Mateos-Díaz ,&nbsp;Jorge A. Rodríguez","doi":"10.1016/j.ab.2025.115769","DOIUrl":"10.1016/j.ab.2025.115769","url":null,"abstract":"<div><div>In this study, we propose a continuous assay that provides a high-throughput, efficient method for screening the regioselectivity of lipases at the <em>sn</em>-1,3 and <em>sn</em>-2 positions on triacylglycerols (TAGs). This assay measures the specific hydrolysis rates at the primary and secondary positions of TAGs derivates containing oleic (O) and punicic (P) acids. The method is based on the absorbance ratio of released punicic acid from the hydrolysis of <em>sn</em>-POP (<em>sn</em>-1,3 regiospecific lipases) and sn-OPO (<em>sn</em>-2 regiospecific lipases). The method was validated using pure lipases with known and unknown regioselectivity. Unexpectedly, we found that recombinant Lipase 4 from <em>Candida rugosa</em> (rCRLip4) exhibited significant <em>sn</em>-2 regioselectivity, indicating greater regioselectivity than recombinant lipase A from <em>Candida antarctica</em> (rCALA). <em>In silico</em> analysis and molecular docking studies were conducted to elucidate the main structural differences between CRLip4 and the non-regioselective isoform CRLip1. This continuous regioselective lipase assay on TAGs is versatile and can be used to screen for <em>sn</em>-1,3, <em>sn</em>-2 or non-regioselective lipases. It holds significant potential applications in the biocatalytic production of structured lipids and other industrial processes where regioselectivity is crucial.</div></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":"700 ","pages":"Article 115769"},"PeriodicalIF":2.6,"publicationDate":"2025-01-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142977147","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A highly sensitive creatine kinase detection in human serum using 11-mercaptoundecanoic acid modified ITO-PET electrodes","authors":"Burçak Demirbakan","doi":"10.1016/j.ab.2025.115768","DOIUrl":"10.1016/j.ab.2025.115768","url":null,"abstract":"<div><div>The enzyme creatine kinase (CK) is a biomarker that plays an extremely significant role in the early detection of cardiovascular disorders. Serum levels of CK are regularly monitored in patients with heart attacks, one of the most critical cardiovascular illnesses. In this study, a highly sensitive electrochemical immunosensor system was designed for the importance of early diagnosis of CK. This immunosensor system was developed by immobilizing 11- mercaptoundecanoic acid (11-MuA) on disposable indium tin oxide–polyethylene terephthalate (ITO-PET) electrodes. Electrochemical impedance spectroscopy (EIS), cyclic voltammetry (CV) and single frequency impedance (SFI) techniques were utilized throughout the immobilization process during the construction of the immunosensor. In addition, the proposed CK immunosensor system involves thorough analytical research, which may include linear determination range, repeatability, reproducibility, square wave voltammetry, storage capability, and regeneration. The suggested immunosensor was also characterized using scanning electron microscopy (SEM). The proposed immunosensor system demonstrated a broad dynamic range (0.1 pg/mL – 100 pg/mL), as well as a low limit of detection (LOD) and a low limit of quantification (LOQ) of 0.018 pg/mL and 0.0394 pg/mL, respectively. Finally, the immunosensor was tested on human serum samples, proving that it could be utilized in clinical situations.</div></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":"700 ","pages":"Article 115768"},"PeriodicalIF":2.6,"publicationDate":"2025-01-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142969401","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Green fabricated bimetallic zinc ferrite nanoparticles mitigate oxidative stress-induced pathogenesis","authors":"Shivakumar Venkataramaiah ,&nbsp;Manjula M. Venkatappa ,&nbsp;Rajesh Rangappa ,&nbsp;Chikkappa Udagani ,&nbsp;Devaraja Sannaningaiah","doi":"10.1016/j.ab.2025.115767","DOIUrl":"10.1016/j.ab.2025.115767","url":null,"abstract":"<div><div>Current study evaluates the beneficial role of bio-functionalized zinc ferrite nanoparticles fabricated from an aqueous extract of <em>Decalepis hamiltonii</em> leaves (<em>DHLE.ZnFe</em>_<em>2</em><em>O</em>_<em>4</em> NPs) on sodium nitrite (NaNO₂) and Diclofenac (DFC) induced oxidative stress in RBCs and Sprague Dawley male rat models. <em>DHLE.ZnFe</em>_<em>2</em><em>O</em>_<em>4</em> NPs were characterized using <em>PXRD, FTIR, SEM-EDAX, HR-TEM</em> and <em>VSM</em>. The data suggests that, <em>DHLE.ZnFe</em>_<em>2</em><em>O</em>_<em>4</em> NPs were crystalline, ellipsoidal in shape with an average size of 10.95 nm and super paramagnetic in nature. <em>DHLE.ZnFe</em>_<em>2</em><em>O</em>_<em>4</em> NPs exhibited anti-oxidant properties by scavenging DPPH, H₂O₂ and reducing ferric to ferrous ions. Furthermore, <em>DHLE.ZnFe</em>_<em>2</em><em>O</em>_<em>4</em> NPs normalized key parameters of oxidative stress such as <em>LPO</em>, <em>PCC</em>, <em>TT</em> and anti-oxidant enzymes (<em>SOD</em> &amp; <em>CAT</em>). Similar to the previous <em>in-vitro</em> results, <em>DHLE.ZnFe</em>_<em>2</em><em>O</em>_<em>4</em> NPs restored all the said stress parameters in homogenates of the liver, kidney, pancreas and heart. In addition, <em>DHLE.ZnFe</em>_<em>2</em><em>O</em>_<em>4</em> NPs repaired Diclofenac induced tissue damage in the liver, kidney, pancreas and heart by regulating all biochemical parameters. Most importantly, <em>DHLE.ZnFe</em>_<em>2</em><em>O</em>_<em>4</em> NPs exhibited anti-inflammatory, anti-diabetic, anti-thrombotic activities and were non-toxic to RBCs. In conclusion, <em>DHLE.ZnFe</em>_<em>2</em><em>O</em>_<em>4</em> NPs through its anti-oxidant potential ameliorate oxidative stress induced pathogenesis such as, inflammation, tissue damage, diabetes and thrombosis.</div></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":"700 ","pages":"Article 115767"},"PeriodicalIF":2.6,"publicationDate":"2025-01-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142942813","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Hydrodynamic characterization of the FtsZ protein from Escherichia coli demonstrates the presence of linear and lateral trimers","authors":"Nelson A. Araujo ,&nbsp;Marcelo Veloso ,&nbsp;Luis Pouchucq","doi":"10.1016/j.ab.2025.115766","DOIUrl":"10.1016/j.ab.2025.115766","url":null,"abstract":"<div><div>FtsZ is a bacterial protein that plays a crucial role in cytokinesis by forming the Z-ring. This ring acts as a scaffold to recruit other division proteins and guide the synthesis of septal peptidoglycan, which leads to cell constriction. In its native state, the FtsZ protein from <em>Escherichia coli</em> (EcFtsZ) is a multi-oligomer comprising dimers, trimers, tetramers, and hexamers in a dynamic self-association equilibrium depending on its concentration. This study employed classical methods of analytical biochemistry that included native polyacrylamide gel electrophoresis, size-exclusion chromatography, sedimentation through sucrose gradients, and chemical cross-linking with formaldehyde to characterize the EcFtsZ. The dimers, trimers, and tetramers are the most prevalent oligomers of the EcFtsZ protein; however, the trimer has been understudied compared to the dimer. In this study, we characterized uncross-linked trimers by exclusion chromatography and crosslinked trimers by sedimentation. The results of size-exclusion chromatography demonstrated that the uncross-linked trimer of EcFtsZ has a mass of 128.8 kDa and a frictional ratio f/f_o of 1.96, which coincides with the theoretical frictional ratio of 1.80 for a linear trimer. The EcFtsZ protein treated with formaldehyde resulted in a polypeptide band of 128 kDa recognized by anti-FtsZ antibodies and a frictional ratio S_max/S_20,w equal to 1.95, which agrees with the theoretical calculation of the frictional ratio of a lateral trimer. The protein-protein interaction prediction program (PEPPI) identified a contact site between subunits in the C-terminal linker region of the EcFtsZ protein, which has the potential to interfere with the recognition of the C-terminal linker by the ClpX(P) protease.</div></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":"699 ","pages":"Article 115766"},"PeriodicalIF":2.6,"publicationDate":"2025-01-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142942815","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}