Journal of the American Society for Mass Spectrometry最新文献_第4页

Characterizing Protein Solvent Accessible Surface Area in Solution by Dual Polarity Native Mass Spectrometry. 双极性天然质谱法表征蛋白质溶液中溶剂可达表面积。

IF 3.1 2区化学

Journal of the American Society for Mass Spectrometry Pub Date : 2025-05-07 Epub Date: 2025-04-01 DOI: 10.1021/jasms.4c00465

Lei Yang, Yi Zhao, Xinyan Fu, Wenjing Zhang, Wei Xu

{"title":"Characterizing Protein Solvent Accessible Surface Area in Solution by Dual Polarity Native Mass Spectrometry.","authors":"Lei Yang, Yi Zhao, Xinyan Fu, Wenjing Zhang, Wei Xu","doi":"10.1021/jasms.4c00465","DOIUrl":"10.1021/jasms.4c00465","url":null,"abstract":"Native mass spectrometry (nMS) is rapidly emerging as a pivotal technique for exploring protein conformations and protein-ligand interactions. Pioneering research has demonstrated that the charge state distribution (CSD) of proteins in native mass spectra can be indicative of their solvent accessible surface area (SASA). Moreover, beyond SASA, it is postulated that the abundance of acidic and basic amino acids on the protein surface may also impact the CSD. Specifically, basic amino acids tend to acquire positive charges during electrospray ionization (ESI), whereas acidic amino acids are prone to adopting negative charges. Consequently, this study investigates the CSDs of globular proteins in both positive and negative ion modes to provide a comprehensive characterization of protein SASA. Experiments were conducted under both native ESI and native nano-ESI conditions. By harnessing the average charges observed across dual polarity nMS data, we achieved significantly enhanced log linear correlations between protein SASA and its CSDs. The coefficient of determination (R2) improved from 0.9866 to 0.9888 under ESI conditions and from 0.9677 to 0.9902 under nano-ESI conditions when compared to models utilizing only positive ion mode data. These findings suggest that the SASA of globular proteins can be effectively characterized through the CSDs derived from dual polarity nMS analysis.","PeriodicalId":672,"journal":{"name":"Journal of the American Society for Mass Spectrometry","volume":" ","pages":"991-998"},"PeriodicalIF":3.1,"publicationDate":"2025-05-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143762577","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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

Discovery and Characterization of Novel G0 Glycan Isomers in an Afucosylated Therapeutic Antibody Using Liquid Chromatography-Mass Spectrometry. 用液相色谱-质谱法发现和鉴定A聚焦治疗性抗体中新的G0聚糖异构体。

IF 3.1 2区化学

Journal of the American Society for Mass Spectrometry Pub Date : 2025-05-07 Epub Date: 2025-03-31 DOI: 10.1021/jasms.4c00474

Miyang Li, Sean Shen, Simon Letarte, Tawnya Flick

{"title":"Discovery and Characterization of Novel G0 Glycan Isomers in an Afucosylated Therapeutic Antibody Using Liquid Chromatography-Mass Spectrometry.","authors":"Miyang Li, Sean Shen, Simon Letarte, Tawnya Flick","doi":"10.1021/jasms.4c00474","DOIUrl":"10.1021/jasms.4c00474","url":null,"abstract":"Therapeutic antibodies are a class of glycoproteins that commonly carry conserved N-glycans at the Fc domain, and the attached N-glycans play a pivotal role in their biological function and efficacy. In this study, we conducted a detailed N-glycan profiling of an afucosylated monoclonal antibody using the Waters GlycoWorks RapiFluor-MS kit and hydrophilic interaction liquid chromatography coupled with fluorescence detection and mass spectrometry (HILIC-FLD-MS). We discovered and reported for the first time novel G0 glycan isomers on a monoclonal antibody. The G0 glycan has a composition of two additional N-acetylglucosamine (GlcNAc) units in addition to the core pentasaccharide structure of N-glycans. The MS/MS profile revealed few fragmentation differences for RapiFluor-MS-labeled glycan isomers. To enhance structural elucidation, a reduction and permethylation assay was performed. Reversed-phase liquid chromatography (RP-LC) separated permethylated glycans due to their altered hydrophobic properties and revealed the presence of additional isomers. The fragmentation of sodium adducts of the permethylated glycans provided distinct patterns among isomers, indicating a bisecting structure for the novel G0 glycan isomers previously identified. Since the bisecting glycans possess one GlcNAc on mannose with 1-4 linkage (bisecting GlcNAc), the other GlcNAc could occupy either branching antenna, resulting in the additional subtle positional isomers, which agrees with our observation. This study underscores the utility of permethylation coupled with advanced chromatography and mass spectrometry techniques to resolve glycan isomers and contributes to a deeper understanding of glycan structural diversity in biologic therapeutic development.","PeriodicalId":672,"journal":{"name":"Journal of the American Society for Mass Spectrometry","volume":" ","pages":"1034-1040"},"PeriodicalIF":3.1,"publicationDate":"2025-05-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143750545","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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

Size-Exclusion Chromatography-Electrospray-Ionization Mass Spectrometry To Characterize End Group and Chemical Distribution of Poly(lactide-co-glycolide) Copolymers. 尺寸排除色谱-电喷雾-电离质谱法表征聚丙交酯-共聚物的端基和化学分布。

IF 3.1 2区化学

Journal of the American Society for Mass Spectrometry Pub Date : 2025-05-07 Epub Date: 2025-03-31 DOI: 10.1021/jasms.4c00447

Masashi Serizawa, Pieter van Delft, Peter J Schoenmakers, Ron A H Peters, Andrea F G Gargano

{"title":"Size-Exclusion Chromatography-Electrospray-Ionization Mass Spectrometry To Characterize End Group and Chemical Distribution of Poly(lactide-co-glycolide) Copolymers.","authors":"Masashi Serizawa, Pieter van Delft, Peter J Schoenmakers, Ron A H Peters, Andrea F G Gargano","doi":"10.1021/jasms.4c00447","DOIUrl":"10.1021/jasms.4c00447","url":null,"abstract":"The characterization of the microstructure of in vivo degradable polyesters is gaining increased interest thanks to their high-performance applications, such as drug delivery systems. The design of such material requires a high level of understanding of the critical material attributes of the polyesters, such as molecular-weight distribution (MWD), chemical-composition distribution (CCD), and end-groups (functionality-type distribution, FTD). Size-exclusion chromatography (SEC) hyphenated with mass spectrometry (MS) is an effective method for analyzing the microstructure of polymers. While the MWD can be determined by size-exclusion chromatography hyphenated with ultraviolet spectrometry and refractive index, the CCD and FTD can be determined by SEC-MS. However, previous applications of SEC-MS have not assessed if polymer fragmentation can occur during the analysis process. In order to correctly interpret CCD and FTD, it is important to establish whether SEC-MS methods can be applied to biodegradable polymers and to recognize if fragmentation processes occur. In this study, we investigate whether SEC-MS methods can be applied to PLGA biodegradable polyesters. The research demonstrates that the choice of alkali metal salt used during ionization can influence the stability of PLGA during SEC-MS analysis. CsI was found to minimize fragmentations during ESI-MS, simplifying the MS spectra and allowing isomeric PLGA structures to be distinguished. The resulting method facilitates FTD and CCD determination. Additionally, when combined with selective degradation, the described method can provide insights into the \"blockiness\" of the polymer and support the development of sequence-controlled PLGA synthesis.","PeriodicalId":672,"journal":{"name":"Journal of the American Society for Mass Spectrometry","volume":" ","pages":"980-990"},"PeriodicalIF":3.1,"publicationDate":"2025-05-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12063183/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143750605","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Enthalpies and Entropies of Activation for sn-1/sn-2 Acyl Chain Loss in Glycerophospholipid Anions via Dipolar DC Kinetics. 偶极直流电动力学对甘油磷脂阴离子中sn-1/sn-2酰基链损失的激活焓和熵。

IF 3.1 2区化学

Journal of the American Society for Mass Spectrometry Pub Date : 2025-05-07 Epub Date: 2025-04-02 DOI: 10.1021/jasms.4c00477

Samantha A Mehnert, Katherine J Lee, Scott A McLuckey

{"title":"Enthalpies and Entropies of Activation for sn-1/sn-2 Acyl Chain Loss in Glycerophospholipid Anions via Dipolar DC Kinetics.","authors":"Samantha A Mehnert, Katherine J Lee, Scott A McLuckey","doi":"10.1021/jasms.4c00477","DOIUrl":"10.1021/jasms.4c00477","url":null,"abstract":"Glycerophospholipids (GPs) have been observed to prefer losing a particular fatty acyl chain over the other, with the preference for the chain in either the sn-1 or sn-2 position being different between various GP classes. It has been assumed that the sn preference results from the entropic favorability of the transition state conformation; however, this has not been measured previously. Here, we demonstrate the application of our previously established collision-based activation method to GP fragmentation. The method utilizes a dipolar direct current (DDC) potential across a pair of opposing rods to force ions out of the center of the ion trap, causing them to undergo radio frequency (RF) heating by absorbing power from the trapping RF field. We confirmed that the previous assumption holds for some species studied here, wherein the ΔH‡ values were nearly identical and the ΔS‡ values showed greater differences between the sn positions. However, some species and ion types seem to be more driven by ΔH‡. Additionally, the loss of the fatty acyl chains as neutrals rather than ions should also be considered if one is to thoroughly weigh which chain is indeed the preferred loss, as including all forms of acyl chain loss results in an overall favorability for the acyl chain in the sn-2 position to be lost. The driving force behind these different losses seems to be a mixture of entropic and enthalpic reasons, with the identity and presence of the headgroup playing an important role in the observed fragmentation and the measured activation parameters.","PeriodicalId":672,"journal":{"name":"Journal of the American Society for Mass Spectrometry","volume":" ","pages":"1041-1051"},"PeriodicalIF":3.1,"publicationDate":"2025-05-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143770899","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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

Native Top-Down Mass Spectrometry Characterization of Model Integral Membrane Protein Bacteriorhodopsin. 模型积分膜蛋白细菌视紫红质的天然自顶向下质谱表征。

IF 3.1 2区化学

Journal of the American Society for Mass Spectrometry Pub Date : 2025-05-07 Epub Date: 2025-04-15 DOI: 10.1021/jasms.4c00439

Jessie Le, Joseph A Loo

{"title":"Native Top-Down Mass Spectrometry Characterization of Model Integral Membrane Protein Bacteriorhodopsin.","authors":"Jessie Le, Joseph A Loo","doi":"10.1021/jasms.4c00439","DOIUrl":"https://doi.org/10.1021/jasms.4c00439","url":null,"abstract":"Bacteriorhodopsin (bR) from Halobacterium salinarum has been a model system for structural biology and is a structural template for the characterization of membrane G-protein couple receptors (GPCRs) in particular. In this study, wild-type bacteriorhodopsin and two single-residue mutants were characterized by native top-down mass spectrometry (nTD-MS) with Orbitrap-based high-energy collision dissociation (HCD) and electron capture dissociation (ECD). After in-source dissociation ejected the membrane protein from detergent micelles, high-resolution native MS measurement allowed for identification of multiple proteoforms as well as lipid-bound forms. Further top-down MS measurements by HCD produced a large number of product ions for in-depth sequencing and unambiguous localization of post-translational modifications. For the first time, native TD-MS with ECD was used to characterize an integral membrane protein. ECD yielded fragments originating from all helices and loop regions, even accessing a sequence stretch that HCD could not. Combining HCD and ECD fragmentation patterns significantly enhanced the sequence coverage of bR. We propose bR to be a model analyte for testing nTD-MS performance for membrane proteins.","PeriodicalId":672,"journal":{"name":"Journal of the American Society for Mass Spectrometry","volume":"36 5","pages":"961-968"},"PeriodicalIF":3.1,"publicationDate":"2025-05-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143954959","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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

Highly Sensitive Chemical Ionization Tandem Mass Spectrometry Method for the Identification of Unsaturated Fatty Acids Derivatized by Dimethyl Disulfide. 高灵敏度化学电离串联质谱法鉴定二甲基二硫化衍生的不饱和脂肪酸。

IF 3.1 2区化学

Journal of the American Society for Mass Spectrometry Pub Date : 2025-05-07 Epub Date: 2025-04-21 DOI: 10.1021/jasms.5c00033

Tingxiang Yang, Lerong Qi, Hao Yang, Yihan Xia, Zhen Wang, Dong Hao Wang

{"title":"Highly Sensitive Chemical Ionization Tandem Mass Spectrometry Method for the Identification of Unsaturated Fatty Acids Derivatized by Dimethyl Disulfide.","authors":"Tingxiang Yang, Lerong Qi, Hao Yang, Yihan Xia, Zhen Wang, Dong Hao Wang","doi":"10.1021/jasms.5c00033","DOIUrl":"https://doi.org/10.1021/jasms.5c00033","url":null,"abstract":"Derivatization of unsaturated fatty acids with dimethyl disulfide (DMDS) and analysis by electron ionization mass spectrometry (EIMS) represent a convenient offline method for the identification of double bond positions. However, the presence of overlapping mass spectra from multiple compounds poses significant challenges for spectral interpretation and library matching, leading to ambiguous molecular information and low sensitivity. To overcome the issue, we developed a novel chemical ionization (CI) tandem mass spectrometry method involving the pre-derivatization with DMDS and collisional activation of [M+47]+ ions generated in the chemical ion source. The method provides better specificity to the analysis of targeted fatty acids and does not require any customized devices. Further, a multiple reaction monitoring (MRM) version of the method was designed by screening all the diagnostic ions of possible double bond positional isomers, which significantly boosts the sensitivity. Compared to the traditional EIMS method, the new method exhibits a lower limit of detection (LLOD) that is one-tenth or lower. Employing the new method, unusual isomer 18:2(5Z,8Z) was co-analyzed with 18:2(9Z,12Z), and a novel 20:2(7Z,10Z) was characterized in human sebum. Additionally, 16:2(9Z,12Z), an odd-chain omega-3 polyunsaturated fatty acid (21:5n-3) and polymethylene-interrupted isomers, i.e. 22:2(7Z,13Z) and 22:2(7Z,15Z) were identified in seafood and related products. Our method can be readily applied to any GC instrument equipped with tandem MS and is expected to facilitate the discovery and identification of unknown fatty acids from food, clinical, and environmental sources.","PeriodicalId":672,"journal":{"name":"Journal of the American Society for Mass Spectrometry","volume":"36 5","pages":"1158-1166"},"PeriodicalIF":3.1,"publicationDate":"2025-05-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143956200","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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

HDgraphiX: A Web-Based Tool for Visualization of Hydrogen/Deuterium Exchange Mass Spectrometry Data. hdgraphhix：一个基于网络的氢/氘交换质谱数据可视化工具。

IF 3.1 2区化学

Journal of the American Society for Mass Spectrometry Pub Date : 2025-05-07 DOI: 10.1021/jasms.5c00005

Kent R Vosper, Algirdas Velyvis, Siavash Vahidi

{"title":"HDgraphiX: A Web-Based Tool for Visualization of Hydrogen/Deuterium Exchange Mass Spectrometry Data.","authors":"Kent R Vosper, Algirdas Velyvis, Siavash Vahidi","doi":"10.1021/jasms.5c00005","DOIUrl":"https://doi.org/10.1021/jasms.5c00005","url":null,"abstract":"Hydrogen-deuterium exchange mass spectrometry (HDX-MS) investigates protein structural changes by measuring deuterium incorporation into the protein amide backbone. Due to the richness of information provided on protein conformational dynamics, HDX-MS data can be challenging to visualize effectively. To address this, we have developed HDgraphiX, a web-based tool that visualizes HDX-MS data by processing outputs from several popular analysis software packages including DynamX, HDExaminer, and HDX Workbench. HDgraphiX performs statistical analyses, filters data based on statistical significance, and presents the results as user-friendly publication-quality visualizations, including heatmaps (Chiclet plots), Woods plots, ladder plots, and volcano plots. Users can optionally generate PyMOL and ChimeraX coloring scripts to map deuterium uptake differences onto protein structures. Additionally, HDgraphiX offers numerous advanced options for customizing data processing and plotting without the need for manual data editing. To improve usability, users have the option to download a file containing all input settings, which can be reuploaded alongside HDX-MS data to avoid manual re-entry of custom settings.","PeriodicalId":672,"journal":{"name":"Journal of the American Society for Mass Spectrometry","volume":"36 5","pages":"1200-1203"},"PeriodicalIF":3.1,"publicationDate":"2025-05-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143951850","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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

Functional Testing and Localization of Tyrosine Sulfation in a Trispecific Antibody. 酪氨酸磺酸在三特异性抗体中的功能测试和定位。

IF 3.1 2区化学

Journal of the American Society for Mass Spectrometry Pub Date : 2025-05-07 Epub Date: 2025-03-25 DOI: 10.1021/jasms.4c00432

Armelle Martelet, Valerie Garrigue, Hélène Le Borgne, Claire Borel, Sylvie Alexandre, Ronan Crépin, Bruno Genet, Haichuan Liu, Yuzhuo Zhang, Séverine Clavier

{"title":"Functional Testing and Localization of Tyrosine Sulfation in a Trispecific Antibody.","authors":"Armelle Martelet, Valerie Garrigue, Hélène Le Borgne, Claire Borel, Sylvie Alexandre, Ronan Crépin, Bruno Genet, Haichuan Liu, Yuzhuo Zhang, Séverine Clavier","doi":"10.1021/jasms.4c00432","DOIUrl":"10.1021/jasms.4c00432","url":null,"abstract":"Mass spectrometry (MS) is a tool of choice for the in-depth characterization of new biotherapeutic molecules such as a complex naturally derived trispecific antibody (tsAb) that presents a tyrosine sulfation within the variable domain. Although tyrosine sulfation is an important post-translational modification responsible for strengthening protein-protein interactions, its localization is challenging, as the sulfate group is very labile using conventional positive ion mode fragmentation techniques. In this work, we describe the combination of functional testing and MS-based methods to study the impact of tyrosine sulfation in the tsAb. The presence of sulfation was confirmed by intact mass and peptide mapping analyses. For unambiguous localization of the sulfate group, electron activated dissociation (EAD) MS/MS experiments were employed to generate diagnostic fragments carrying an intact sulfate group. We also demonstrated that a significant decrease in binding of the tsAb to the target antigen was observed following the sulfatase treatment. Taken together, the results from this study support the notion that tyrosine sulfation plays an important role in protein-protein interactions.","PeriodicalId":672,"journal":{"name":"Journal of the American Society for Mass Spectrometry","volume":" ","pages":"952-960"},"PeriodicalIF":3.1,"publicationDate":"2025-05-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143708000","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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

Norm ISWSVR Enhanced Data Repeatability and Accuracy in Large-Scale Targeted Quantification Metabolomics. Norm ISWSVR提高了大规模靶向量化代谢组学数据的可重复性和准确性。

IF 3.1 2区化学

Journal of the American Society for Mass Spectrometry Pub Date : 2025-05-07 Epub Date: 2025-04-03 DOI: 10.1021/jasms.4c00467

Jinpeng Bai, Chenxi Li, Mingmin Qian, Xian Ding, Qian Li, Yanhua Chen, Zhaoying Wang, Zeper Abliz

{"title":"Norm ISWSVR Enhanced Data Repeatability and Accuracy in Large-Scale Targeted Quantification Metabolomics.","authors":"Jinpeng Bai, Chenxi Li, Mingmin Qian, Xian Ding, Qian Li, Yanhua Chen, Zhaoying Wang, Zeper Abliz","doi":"10.1021/jasms.4c00467","DOIUrl":"10.1021/jasms.4c00467","url":null,"abstract":"Targeted quantification metabolomics provides dynamic insights across various domains within the life sciences. Nevertheless, maintaining high-quality data obtained through liquid chromatography-mass spectrometry presents ongoing challenges. It is essential to develop normalization methods to correct for unwanted variations in metabolomic profiling such as batch effects and analytical drift. In this study, we assessed the normalization efficacy of Norm ISWSVR in targeted quantification metabolomics by comparing it with IS normalization and SERRF normalization. Consequently, Norm ISWSVR demonstrated exceptional efficacy in mitigating batch effects and reducing the relative standard deviation of quality control samples, in addition to correcting signal drift. Following normalization with Norm ISWSVR, the number of metabolites suitable for quantification increased with high precision. Collectively, Norm ISWSVR proves to be a robust and reliable method for enhancing data quality in targeted metabolomics, establishing itself as a promising approach for metabolomics research.","PeriodicalId":672,"journal":{"name":"Journal of the American Society for Mass Spectrometry","volume":" ","pages":"1027-1033"},"PeriodicalIF":3.1,"publicationDate":"2025-05-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143778634","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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

Simultaneous Quantitative Detection of Cysteine and Homocysteine Labeled by 1-Pyrenecarboxaldehyde Using MALDI-TOF MS. MALDI-TOF质谱同时定量检测1-芘甲酸标记的半胱氨酸和同型半胱氨酸。

IF 3.1 2区化学

Journal of the American Society for Mass Spectrometry Pub Date : 2025-05-07 Epub Date: 2025-04-15 DOI: 10.1021/jasms.4c00512

Liming Guo, Hao Wang, Liang Song, Chunsheng Xiao, Jiarui Li, Xinhua Guo

{"title":"Simultaneous Quantitative Detection of Cysteine and Homocysteine Labeled by 1-Pyrenecarboxaldehyde Using MALDI-TOF MS.","authors":"Liming Guo, Hao Wang, Liang Song, Chunsheng Xiao, Jiarui Li, Xinhua Guo","doi":"10.1021/jasms.4c00512","DOIUrl":"https://doi.org/10.1021/jasms.4c00512","url":null,"abstract":"Cysteine (Cys) and homocysteine (Hcy) are two important reducing agents in living organisms and play crucial roles in many physiological processes. The quantitative analysis of Cys and Hcy holds significance in exploring the functions of biothiols in biological. In this work, 1-pyrenecarboxaldehyde (1-py) with high derivatization efficiency and ionization efficiency was used for quantitative analysis of Cys and Hcy by matrix-assisted laser desorption and ionization time-of-flight mass spectrometry (MALDI-TOF MS). After 1-py derivatization, the detection limit of Cys and Hcy can reach as low as 250 amol/L. Without internal standards, the simultaneous quantitative detection of Cys and Hcy was achieved by analyzing the proportion of peak intensities of derivative products to total compounds. The linear quantitative ranges for Cys and Hcy were over the concentrations from 5 to 2500 μM. Moreover, the specific hydrogen loss of the derivatized products was observed in MALDI-TOF detection, and the potential fragment pathway and nitrogen protonation mechanism were demonstrated through density functional theory (DFT) calculations. Finally, this method was successfully applied to the quantification of Cys and Hcy in HepG2 cell lysate, offering a rapid and highly sensitive approach for the quantitative analysis of Cys and Hcy using MALDI-TOF MS.","PeriodicalId":672,"journal":{"name":"Journal of the American Society for Mass Spectrometry","volume":"36 5","pages":"1077-1083"},"PeriodicalIF":3.1,"publicationDate":"2025-05-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143956759","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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