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Recent advances in active chromophores for detecting gamma-hydroxybutyric acid (GHB) related illicit drugs
IF 4.2 3区 化学
Analyst Pub Date : 2025-03-31 DOI: 10.1039/d5an00167f
Chun Yang, Hongxian Yang, Zhen Yao, Taihong Liu
{"title":"Recent advances in active chromophores for detecting gamma-hydroxybutyric acid (GHB) related illicit drugs","authors":"Chun Yang, Hongxian Yang, Zhen Yao, Taihong Liu","doi":"10.1039/d5an00167f","DOIUrl":"https://doi.org/10.1039/d5an00167f","url":null,"abstract":"Gamma-hydroxybutyric acid (GHB) and its related illicit drugs are of particular forensic interest due to their evil abuse as recreational drugs and implications in drug-facilitated sexual assault. The rapid and complete metabolism of GHB in body result in a short evidence collection window for the forensic experts, and the challenges in differentiating exogenous addition in spiked drinking and low endogenous levels of GHB exist simultaneously. Consequently, development of real-time and on-site detection strategies for GHB plays vital roles in tackling drug-facilitated crimes. Recently, fluorescent and colorimetric strategies have emerged as promising approaches in this field, offering multiple merits of high sensitivity, specificity, ease of handling, and cumulative signaling effects. This minireview outlines the endogenous levels of GHB in body and possible metabolism pathways, summarizes the recent advances of active chromophores, elucidates the corresponding sensing characteristics, and then exemplifies the developed sensing strips and detection kits based on the optimized chromophores mostly in five years. Additionally, the perspectives of the relevant studies are discussed properly.","PeriodicalId":63,"journal":{"name":"Analyst","volume":"33 1","pages":""},"PeriodicalIF":4.2,"publicationDate":"2025-03-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143736788","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Teicoplanin-cyclodextrin bilayer chiral stationary phase boosts chiral separation of native amino acids
IF 4.2 3区 化学
Analyst Pub Date : 2025-03-28 DOI: 10.1039/d5an00156k
Chenglin Zhang, Xiangyun Yu, Yin Xiao, Qiang Zhang
{"title":"Teicoplanin-cyclodextrin bilayer chiral stationary phase boosts chiral separation of native amino acids","authors":"Chenglin Zhang, Xiangyun Yu, Yin Xiao, Qiang Zhang","doi":"10.1039/d5an00156k","DOIUrl":"https://doi.org/10.1039/d5an00156k","url":null,"abstract":"Separating enantiomers is crucial for advancing scientific research. Teicoplanin-based columns exhibit unique advantages in the chiral separation of native amino acids, and its further improvement for better separation performance is valuable. This work reported double-layer bridged columns, in which different amounts of cyclodextrin (CD) were directly bridged onto teicoplanin (TK) molecules on chiral stationary phase (TK CSP) to synthesize TK-CD-2 CSP and TK-CD-6 CSP. Additionally, a column with the two chiral selectors (TK and CD) arranged in a single layer on the silica gel surface (TK+CD CSP) was synthesized, which served as a control group. The results revealed TK-CD-6 CSP had better chiral separation ability of native amino acids and wider range of separable chiral molecules than TK, TK+CD, TK-CD-2 CSPs. The possible mechanism is the dual-layer structure of the column (TK-CD-6 CSP) effectively reduces the competition between different chiral selectors and provides more chiral recognition sites, which enhances the separation ability and achieves broad-spectrum chiral separation. This work offers valuable inspirations for enhancing the chiral separation ability of teicoplanin-based columns and developing liquid chromatographic strategy with separation diversity.","PeriodicalId":63,"journal":{"name":"Analyst","volume":"30 1","pages":""},"PeriodicalIF":4.2,"publicationDate":"2025-03-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143723277","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
A catalytic hairpin assembly system with sliding replication for the detection of piRNAs
IF 4.2 3区 化学
Analyst Pub Date : 2025-03-28 DOI: 10.1039/d5an00076a
Hui Yang, Feng Gong, Chengjiao Yao, Jiao Cheng, Shuang He, Yijie Jia, Yuxiang Zhang, Qiang Ma, Xiaolan Guo, Hongfei He, Xiaowu Zhong
{"title":"A catalytic hairpin assembly system with sliding replication for the detection of piRNAs","authors":"Hui Yang, Feng Gong, Chengjiao Yao, Jiao Cheng, Shuang He, Yijie Jia, Yuxiang Zhang, Qiang Ma, Xiaolan Guo, Hongfei He, Xiaowu Zhong","doi":"10.1039/d5an00076a","DOIUrl":"https://doi.org/10.1039/d5an00076a","url":null,"abstract":"As novel noncoding small RNA molecules, piRNAs play crucial roles in cancer development. However, due to their short sequences, easy degradation, and low abundance, developing specific detection methods is challenging. Rapid and early detection is important for the early clinical detection of tumours. Here, a novel one-step, dual-signal amplification piRNA detection system based on sliding replication and catalytic hairpin assembly (CHA), termed CTA, was developed for rapid, ultrasensitive and specific detection of piRNA-823. By utilizing the unique characteristics of tandem repeat sequences to improve amplification efficiency and fluorescence signal intensity, CTA achieved efficient target recognition and signal amplification by embedding tandem repeat sequences in one of the hairpin probes and utilizing chain displacement reactions to produce strong and detectable signals. CTA detected piRNA-823 with a low detection limit of 65.31 fM. Moreover, the whole detection process could be completed within 45 min. In addition, CTA performed excellently in the detection of cell and cancer samples, and its detection results were consistent with those of RT‒qPCR. More importantly, CTA was successfully applied to effectively differentiate between normal individuals and patients with colorectal cancer. These findings suggest its promising application in the diagnosis of cancer.","PeriodicalId":63,"journal":{"name":"Analyst","volume":"35 1","pages":""},"PeriodicalIF":4.2,"publicationDate":"2025-03-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143723385","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Recent advances in electrochemical detection of reactive oxygen species: a review
IF 4.2 3区 化学
Analyst Pub Date : 2025-03-28 DOI: 10.1039/d4an01533a
Hamidreza Ghaedamini, Dong-Shik Kim
{"title":"Recent advances in electrochemical detection of reactive oxygen species: a review","authors":"Hamidreza Ghaedamini, Dong-Shik Kim","doi":"10.1039/d4an01533a","DOIUrl":"https://doi.org/10.1039/d4an01533a","url":null,"abstract":"Reactive oxygen species (ROS) are mainly generated as a result of cellular metabolism in plants and animals, playing a crucial role in cellular signaling mechanisms. The excessive generation of ROS leads to oxidative stress, which is associated with numerous diseases such as cancer, diabetes, and neurodegenerative disorders. Superoxide (O<small><sub>2</sub></small>˙<small><sup>−</sup></small>), hydrogen peroxide (H<small><sub>2</sub></small>O<small><sub>2</sub></small>), and hydroxyl radicals (˙OH) are the most common ROS involved in a wide range of human diseases. Therefore, sensitive and selective detection of these ROS is of paramount importance for understanding their roles in biological systems and for disease diagnosis. Among the various detection methods, electrochemical techniques have gained significant attention due to their high sensitivity, selectivity, and real-time monitoring capabilities. Electrochemical methods incorporate both organic and inorganic molecules to detect and monitor ROS, facilitating a deeper understanding of how their levels influence diseases linked to oxidative stress. This review aims to provide a critical discussion on the recent advances in electrochemical methods for detecting O<small><sub>2</sub></small>˙<small><sup>−</sup></small>, H<small><sub>2</sub></small>O<small><sub>2</sub></small>, and ˙OH. The review also highlights the application of these electrochemical techniques in detecting ROS in living cells and discusses the challenges and future perspectives in this field.","PeriodicalId":63,"journal":{"name":"Analyst","volume":"61 1","pages":""},"PeriodicalIF":4.2,"publicationDate":"2025-03-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143723249","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Cationic and anionic phenothiazine derivatives: electrochemical behavior and application in DNA-sensor development
IF 4.2 3区 化学
Analyst Pub Date : 2025-03-27 DOI: 10.1039/d5an00164a
Anastasya N. Malanina, Yury Kuzin, Pavel L. Padnya, Alexey N. Ivanov, Ivan Stoikov, Gennady Evtugyn
{"title":"Cationic and anionic phenothiazine derivatives: electrochemical behavior and application in DNA-sensor development","authors":"Anastasya N. Malanina, Yury Kuzin, Pavel L. Padnya, Alexey N. Ivanov, Ivan Stoikov, Gennady Evtugyn","doi":"10.1039/d5an00164a","DOIUrl":"https://doi.org/10.1039/d5an00164a","url":null,"abstract":"The global shift toward personalized medicine and point-of-care testing drives the need for improved analytical performance in sensor technologies. A critical aspect of developing voltammetric sensors lies in identifying novel materials for electrode modification. This study focuses on the electrochemical investigation of two phenothiazine derivatives with distinct terminal functional groups: a cationic compound, 3,7-bis(4-aminophenylamino)phenothiazin-5-ium chloride, and an anionic compound, 3,7-bis(4-carboxyphenylamino)phenothiazin-5-ium chloride. Cyclic voltammetry, electrochemical impedance spectroscopy, quartz crystal microbalance, and scanning electron microscopy were employed to characterize these novel materials. The mutual influence of phenothiazines on voltammetric signals in solution was analyzed, revealing changes in the number of hydrogen ions transferred when transitioning from an individual cationic derivative solution to a mixed solution with the anionic derivative. Two approaches for modifying glassy carbon electrodes were studied: electropolymerization from a mixture of phenothiazines and consecutive electrodeposition of polymeric films from individual solutions. Morphological and quantitative differences in the resulting electrode films were observed, with the latter method yielding a more uniform and thicker layer of redox-active material. A DNA-sensor based on the consecutive electrodeposited films for the detection of doxorubicin was developed. The redox peak currents of the electropolymerized phenothiazine products exhibited a linear response to the logarithm of doxorubicin concentration. The sensor displayed two distinct linear ranges from 0.1 fM to 1 nM and from 1 nM to 1 μM, with a limit of detection calculated from first range at 0.6 fM. This DNA-sensor offers promising applications for advancing point-of-care diagnostics.","PeriodicalId":63,"journal":{"name":"Analyst","volume":"3 1","pages":""},"PeriodicalIF":4.2,"publicationDate":"2025-03-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143713219","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Sensitive detection of small extracellular vesicles using gold nanostar-based SERS assay
IF 4.2 3区 化学
Analyst Pub Date : 2025-03-27 DOI: 10.1039/d5an00110b
Long Ngo, Anastasiia Tukova, wei zhang, Simon Chang-Hao Tsao, Yuling Wang
{"title":"Sensitive detection of small extracellular vesicles using gold nanostar-based SERS assay","authors":"Long Ngo, Anastasiia Tukova, wei zhang, Simon Chang-Hao Tsao, Yuling Wang","doi":"10.1039/d5an00110b","DOIUrl":"https://doi.org/10.1039/d5an00110b","url":null,"abstract":"Small extracellular vesicles (sEVs) are lipid bilayer-bound vesicles that carry critical biomarkers for disease detection. However, the inherent heterogeneity and complexity of sEVs molecular characteristics pose significant challenges for accurate and comprehensive molecular profiling. Traditional analytical methods, including immunoblotting, enzyme-linked immunosorbent assay (ELISA), and flow cytometry, exhibit several limitations, such as being time-consuming, requiring large sample volumes, and demonstrating relatively low sensitivity. Therefore, there is an urgent need to develop a highly sensitive and specific assay for the reliable detection of sEVs. Surface-enhanced Raman scattering (SERS) assays have emerged as a promising approach for sEV detection, offering advantages including high sensitivity and specificity. In the proposed SERS assay, SERS nanotags – comprising nanoparticles coated with Raman-active molecules and conjugated with antibodies – are employed to label surface-bound molecules on sEVs. This approach facilitates the generation of a high-intensity signal from molecules present in low abundance. Recently, anisotropic nanoparticles, such as star-shaped nanostructures, have garnered interest due to their ability to significantly amplify generated SERS signals for ultra-sensitive biomarkers detection. In this study, we explore the application of gold nanostars (AuNSs) as SERS nanotags for the detection of sEVs. In principle, AuNSs-based SERS nanotags were used to label EpCAM protein, that can be found on the surface of cancer cell derived sEVs, then sEVs labelled SERS nanotags were captured with CD9 conjugated magnetic beads to form immunocomplex, which exhibits SERS signal. Our results demonstrate that the proposed SERS assay utilizing AuNSs provides high specificity and sensitivity, with a detection limit as low as 2.47 × 103 particles/µL. Furthermore, the assay was tested with spiked plasma samples (cancer cell-derived sEVs spiked in healthy plasma), showing that its specificity remains unaffected by the presence of plasma. These findings suggest that the SERS assay incorporating AuNSs holds significant promise as an effective and reliable detection method for potential clinical applications.","PeriodicalId":63,"journal":{"name":"Analyst","volume":"35 1","pages":""},"PeriodicalIF":4.2,"publicationDate":"2025-03-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143713220","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Tyramide Signal Amplification for Highly Sensitive Multiplex Immunoassay based on Encoded Hydrogel Microparticles
IF 4.2 3区 化学
Analyst Pub Date : 2025-03-26 DOI: 10.1039/d5an00078e
Jun Hee Choi, Young Hee Kim, Jiwoo Kim, Yong Jun Lim, Min Jung Kim, Ki Wan Bong
{"title":"Tyramide Signal Amplification for Highly Sensitive Multiplex Immunoassay based on Encoded Hydrogel Microparticles","authors":"Jun Hee Choi, Young Hee Kim, Jiwoo Kim, Yong Jun Lim, Min Jung Kim, Ki Wan Bong","doi":"10.1039/d5an00078e","DOIUrl":"https://doi.org/10.1039/d5an00078e","url":null,"abstract":"Proteins play a crucial role as mediators of immune regulation, homeostasis, and metabolism, making their quantification essential for understanding the disease mechanisms in biomedical research and clinical diagnostics. However, conventional methods have difficulty when detecting proteins in clinical samples in terms of sensitivity, dynamic range, and multiplex capacity. In this study, we develop a highly sensitive multiplex immunoassay based on encoded hydrogel microparticles (MP) utilizing tyramide signal amplification (TSA). The combination of large multiplexing capacity of encoded hydrogel microparticles and the signal amplification of TSA enables highly sensitive multiplex immunoassay. By employing the TSA, we are able to achieve bigger detection signals with higher specificity. We effectively decrease the non-specific bindings in the hydrogel network by blocking the unreacted acrylate double bonds remaining after the capture antibody conjugation step and acquire 3-fold increased signal-to-noise ratio. Also, we optimize three parameters mainly affecting the assay sensitivity: the detection antibody concentration, the biotinyl tyramide concentration and the TSA reaction time. This approach leads to a significant improvement in the assay sensitivity, achieving a limit of detection as low as 58 fg/mL. Compared to the previous method, the assay sensitivity is enhanced 10-fold. In addition, the multiplex capability of the assay is validated by detecting cytokines IL-4, IL-5, IL-6, IL-9, and IL-17, with no observed cross-reactivity. Finally, with enhanced sensitivity, we demonstrate the clinical applicability of our platform by successfully multiplexing these cytokines at concentrations down to several hundreds of fg/mL within the human serum, which could not be detected using previous methods.","PeriodicalId":63,"journal":{"name":"Analyst","volume":"108 1","pages":""},"PeriodicalIF":4.2,"publicationDate":"2025-03-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143702960","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Paper and Cloth-Based Microfluidic Chip for Rapid Cysteine Detection in Deep-Sea Cold Seep
IF 4.2 3区 化学
Analyst Pub Date : 2025-03-26 DOI: 10.1039/d5an00109a
Die Hu, Jiawen Xiang, Jingying Guo, Chao Wang, Ji Qi, Bowei Li, Xiaoyan Wang, Xin Zhang, Lingxin Chen, Xuming Zhuang
{"title":"Paper and Cloth-Based Microfluidic Chip for Rapid Cysteine Detection in Deep-Sea Cold Seep","authors":"Die Hu, Jiawen Xiang, Jingying Guo, Chao Wang, Ji Qi, Bowei Li, Xiaoyan Wang, Xin Zhang, Lingxin Chen, Xuming Zhuang","doi":"10.1039/d5an00109a","DOIUrl":"https://doi.org/10.1039/d5an00109a","url":null,"abstract":"In this study, we developed a simple and cost-effective paper and cloth-based microfluidic fluorescence sensing device for the selective and quantitative detection of L-cysteine (L-Cys) in deep-sea water, addressing the need for efficient monitoring of this critical metabolite of deep-sea creature’s biomarker. The device employs gold nanoparticles (Au NPs) immobilized on a cloth-based substrate and Rhodamine B (Rh B) molecules. In the presence of L-Cys, strong interactions between L-Cys and the Au NPs release Rh B molecules, restoring fluorescence proportional to the cysteine concentration. The device achieves a low detection limit of 0.52 nM with a dynamic range of 1-100 nM. It demonstrates excellent selectivity over other amino acids, stability over 30 days, and reproducibility. Its practical applicability was confirmed using deep-sea cold seep water samples, yielding recoveries of 98.07%-102.62%. Compared to existing methods, this platform offers enhanced portability, lower cost, and faster response, making it suitable for in-situ environmental monitoring.","PeriodicalId":63,"journal":{"name":"Analyst","volume":"61 1","pages":""},"PeriodicalIF":4.2,"publicationDate":"2025-03-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143702962","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
A DNA concatemer-encoded CRISPR/Cas12a fluorescence sensor for sensitive detection of Pb2+ based on DNAzymes
IF 4.2 3区 化学
Analyst Pub Date : 2025-03-26 DOI: 10.1039/d5an00189g
Shaoying He, Wei Lin, Xin Liu, Fei Li, Hong Liang, Huo Xu, Chunhua Lu, Chao Xing
{"title":"A DNA concatemer-encoded CRISPR/Cas12a fluorescence sensor for sensitive detection of Pb2+ based on DNAzymes","authors":"Shaoying He, Wei Lin, Xin Liu, Fei Li, Hong Liang, Huo Xu, Chunhua Lu, Chao Xing","doi":"10.1039/d5an00189g","DOIUrl":"https://doi.org/10.1039/d5an00189g","url":null,"abstract":"Lead pollution presents a significant threat to ecological systems and human health, underscoring the urgent need for highly sensitive detection methods. Herein, we introduce a novel DNA concatemer-encoded CRISPR/Cas12a fluorescence sensor (MDD-Cas12a) for sensitive detection of Pb<small><sup>2+</sup></small> based on DNAzymes. To accomplish this, we designed a substrate strand containing a long DNA concatemer encoding multiple protospacer adjacent motifs (PAMs) and protospacer sequences for activation of the CRISPR/Cas12a system. The DNA concatemer was subsequently anchored to the surface of magnetic beads (MBs) to fabricate a MBs-DNA concatemer nanoprobe. In the presence of Pb<small><sup>2+</sup></small>, the DNAzyme structure catalyzes the cleavage of the substrate strand, leading to the release of DNA concatemers. Following magnetic separation, the released DNA concatemers significantly activate the non-specific trans-cleavage activity of the Cas12a/crRNA complex. The fluorescence reporter DNA is then completely cleaved by the activated Cas12a/crRNA complex, and the Pb<small><sup>2+</sup></small> concentration in the sample can be quantified by measuring the fluorescence signal. By harnessing the specific recognition capability of DNAzymes for Pb<small><sup>2+</sup></small>, the programmability of DNA concatemers, and the self-amplification features of the CRISPR/Cas12a system, the MDD-Cas12a platform demonstrates high sensitivity and specificity for detecting Pb<small><sup>2+</sup></small> in milk and lake water samples.","PeriodicalId":63,"journal":{"name":"Analyst","volume":"68 1","pages":""},"PeriodicalIF":4.2,"publicationDate":"2025-03-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143702966","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Development of a multiple reaction monitoring (MRM)-based LC-MS/MS method for the quantification of post-translational modifications on histone H3 variants in Arabidopsis thaliana
IF 4.2 3区 化学
Analyst Pub Date : 2025-03-25 DOI: 10.1039/d4an01563k
Yajun Hu, Chenxi He, Lei Zhang, Hong Jin
{"title":"Development of a multiple reaction monitoring (MRM)-based LC-MS/MS method for the quantification of post-translational modifications on histone H3 variants in Arabidopsis thaliana","authors":"Yajun Hu, Chenxi He, Lei Zhang, Hong Jin","doi":"10.1039/d4an01563k","DOIUrl":"https://doi.org/10.1039/d4an01563k","url":null,"abstract":"<em>Background</em>: although the canonical histone H3.1 and its variant H3.3 differ by only four amino acids, they exhibit distinct genome-wide binding patterns and regulate different biological pathways. Post-translational modifications (PTMs) on histone tails mediate diverse downstream regulatory processes, raising the question of whether H3.1 and H3.3 harbor variant-specific modifications. However, the minimal amino acid differences between H3.1 and H3.3 make it challenging to distinguish and quantify them using traditional methods. <em>Results</em>: in this study, we developed an integrated multiple reaction monitoring (MRM)-based LC-MS/MS method to accurately differentiate and quantify K27 and K36 modifications on H3.1 and H3.3 in <em>Arabidopsis thaliana</em>. Our findings show that H3.1 contains more K27 methylation marks, associated with gene silencing, whereas H3.3 is enriched in K36 methylation, a mark of active transcription. Additionally, we compared K36 methylation levels in wild-type and SDG8-depleted cells, revealing that the K36 methyltransferase SDG8 shows a strong preference for H3.3 in both <em>in vitro</em> and <em>in vivo</em> assays. By analyzing public datasets, we further identified a strong correlation between H3.3 and the regions where H3K36me3 levels were reduced in <em>sdg8</em> knockout cells. <em>Significance</em>: the MRM-based LC-MS/MS method established in this study provides a reliable and robust tool for the quantification of histone H3.1 and H3.3 PTMs in <em>Arabidopsis thaliana</em>. We demonstrate that the methyltransferase SDG8 shows a strong substrate preference for H3.3. This discovery highlights the importance of histone variant-specific modifications and suggests new avenues for research into their regulatory roles.","PeriodicalId":63,"journal":{"name":"Analyst","volume":"32 1","pages":""},"PeriodicalIF":4.2,"publicationDate":"2025-03-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143695769","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
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