Journal of the American Society for Mass Spectrometry最新文献

{"title":"Novel Immuno-Immobilization Mass-Spectrometry-Based Assay to Enable Structure-Activity Relationship Development of Emopamil Binding Protein (EBP) Inhibitors.","authors":"Ronghai Cheng, Paramasivam Murugan, Meiyi Zhang, Daniel Tekverk, Wanru Li, Katrina Ngyuen, Aram Chang, Rajesh Kumar, Karen S Conrad, Howard Jones, Martin Himmelbauer, Reza Nemati","doi":"10.1021/jasms.5c00121","DOIUrl":"https://doi.org/10.1021/jasms.5c00121","url":null,"abstract":"<p><p>Emopamil binding protein (EBP) is a transmembrane isomerase that catalyzes the conversion of 8,9-unsaturated sterols to 7,8-unsaturated sterols in the cholesterol biosynthesis pathway. Accumulation of 8,9-unsaturated sterols promotes oligodendrocyte progenitor cell (OPC) differentiation into new myelinating oligodendrocytes, making EBP inhibition a promising therapeutic strategy for neurodegenerative diseases such as multiple sclerosis. Structure-activity relationship (SAR)-driving assays are critical for advancing drug candidates through preclinical drug discovery. However, EBP is a challenging target due to low production yield and loss of enzymatic activity during the purification process. In addition, assay throughput is limited by chromatographic separation of EBP isomers. Here, we describe a robust functional assay to support SAR of EBP small molecule inhibitors. Immuno-mobilization of the active form of EBP, overexpressed in Expi293F cells, was coupled with liquid chromatography-atmospheric pressure chemical ionization multiple reaction monitoring MS (LC-APCI MRM MS) to quantify the potency of EBP inhibitors. This approach significantly enhanced EBP enzymatic activity at nanomolar concentrations compared to that of purified recombinant EBP (recEBP). Furthermore, automating sterol extraction and integrating a dual-arm high-speed LC autosampler further improved assay throughput. This approach showed excellent reproducibility and robustness suitable for SAR development. This was evidenced by high-quality individual concentration-response curves (CRCs) with confidence limit ratios (CLRs) below 5; a normal distribution of reference EBP inhibitor pIC₅₀ values with a two-standard deviation (2-SD) of 0.61 based on 72 independent measurements; and consistent IC₅₀ values for over 20 active EBP inhibitors spanning a broad potency range across two independent assay runs, with a mean pIC₅₀ difference between runs of -0.05 and a 2-SD interval of 0.24.</p>","PeriodicalId":672,"journal":{"name":"Journal of the American Society for Mass Spectrometry","volume":" ","pages":""},"PeriodicalIF":3.1,"publicationDate":"2025-07-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144551623","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}

{"title":"","authors":"Sarah R. Zelle,&nbsp;W. Hayes McDonald,&nbsp;Kristie L. Rose,&nbsp;Hassane S. Mchaourab and Kevin L. Schey*,&nbsp;","doi":"","DOIUrl":"","url":null,"abstract":"","PeriodicalId":672,"journal":{"name":"Journal of the American Society for Mass Spectrometry","volume":"36 7","pages":"XXX-XXX 962–968"},"PeriodicalIF":3.1,"publicationDate":"2025-07-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.acs.org/doi/pdf/10.1021/jasms.5c00087","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144521933","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}

{"title":"","authors":"Weiwen Sun,&nbsp;Xiaotong Geng,&nbsp;Jihong Xu,&nbsp;Yong Huang,&nbsp;Shifa Zhong*,&nbsp;Lushi Lian* and Xiaohong Guan,&nbsp;","doi":"","DOIUrl":"","url":null,"abstract":"","PeriodicalId":672,"journal":{"name":"Journal of the American Society for Mass Spectrometry","volume":"36 7","pages":"XXX-XXX 962–968"},"PeriodicalIF":3.1,"publicationDate":"2025-07-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.acs.org/doi/pdf/10.1021/jasms.5c00088","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144521934","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}

{"title":"Advancing Stable Isotope Analysis with Orbitrap-MS for Fatty Acid Methyl Esters and Complex Lipid Matrices.","authors":"Gabriel F Dos Santos, Giovanni B Bevilaqua, Alexis Gilbert, Hugo G Machado, Maxime Julien, Gesiane S Lima, Nerilson M Lima, Júlio C O Ribeiro, Alexandre A Ferreira, Ygor S Rocha, Boniek Gontijo","doi":"10.1021/jasms.5c00092","DOIUrl":"10.1021/jasms.5c00092","url":null,"abstract":"<p><p>Isotopic analysis plays a crucial role in different scientific fields, offering valuable insights that aid in elucidating biosynthetic pathways, determining geographic origin, and identifying product adulteration. Established mass spectrometry techniques for isotopic analysis require the conversion of samples into gases prior to introduction into the systems. Moreover, the ionization process in these methods is destructive, potentially leading to the loss of essential molecular structure information. Thus, alternative analytical methods, such as Orbitrap-MS, could be a useful tool to determine stable isotope ratios. This paper describes an Orbitrap-based method using stearic acid methyl ester as a model molecule to determine the stable isotopic ratios of fatty acids and fatty acid methyl esters (FAMEs) in different vegetable butters. Orbitrap analyses were performed in positive ionization mode with both [M + H]⁺ and [M + Na]⁺ ions considered for the analysis. Nine standards (Std 1-Std 9) and three vegetable butters (cupuaçu, cocoa, and shea) were employed in the study. The standards were employed to develop the method and were measured using HPLC and a dual-inlet system. Both injections achieved high precision (<1.5‰) when compared with the IRMS data; however, the HPLC showed the most accuracy and was selected for direct injection measurement of the natural samples. Our results demonstrated the efficiency of the ESI-Orbitrap system in differentiating sources based on δ¹³C values. This study not only advances the use of high-resolution mass spectrometry for isotope analysis but also opens new avenues for applying stable isotopes in food sciences.</p>","PeriodicalId":672,"journal":{"name":"Journal of the American Society for Mass Spectrometry","volume":" ","pages":"1527-1535"},"PeriodicalIF":3.1,"publicationDate":"2025-07-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144315736","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}

{"title":"","authors":"Ning Ding,&nbsp; and ,&nbsp;Robert J. Levis*,&nbsp;","doi":"","DOIUrl":"","url":null,"abstract":"","PeriodicalId":672,"journal":{"name":"Journal of the American Society for Mass Spectrometry","volume":"36 7","pages":"XXX-XXX 962–968"},"PeriodicalIF":3.1,"publicationDate":"2025-07-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.acs.org/doi/pdf/10.1021/jasms.5c00094","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144521936","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}

{"title":"","authors":"Yuetian Yan,&nbsp;Tao Xing,&nbsp;Shunhai Wang* and Ning Li,&nbsp;","doi":"","DOIUrl":"","url":null,"abstract":"","PeriodicalId":672,"journal":{"name":"Journal of the American Society for Mass Spectrometry","volume":"36 7","pages":"XXX-XXX 962–968"},"PeriodicalIF":3.1,"publicationDate":"2025-07-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.acs.org/doi/pdf/10.1021/jasms.5c00089","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144521940","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}

{"title":"Key Lipid Reprogramming Revealed in Gastric Signet Ring Cell Carcinoma by Spatial Mass Spectrometry Metabolomics.","authors":"Yuying Shi, Qi Yang, Botao Xu, Chuang Yuan, Keke Qi, Yang Pan, Zhe Wang, Qitao Chen, Dong Sun, Weizhu Zhao, Tao Zhang, Chengyuan Liu, Cheng Wang, Jie Chai","doi":"10.1021/jasms.4c00505","DOIUrl":"https://doi.org/10.1021/jasms.4c00505","url":null,"abstract":"<p><p>Gastric signet ring cell carcinoma (GSRC) is an aggressive subtype of gastric cancer (GC) with a poor prognosis. The lack of a systematic molecular and metabolic heterogeneity overview has led to slow progress in clinical practice. This study used mass spectrometry imaging (MSI) to investigate the metabolic landscape of GSRC in GC tissue with various differentiation grades. Our comprehensive spatial profiling of metabolites and lipids unveiled distinct metabolic signatures across different tissue subregions. A substantial number of lipidomic biomarkers associated with GSRC were identified, including phosphatidylethanolamine <i>N</i>-methyl (PE-NMe), phosphatidylethanolamine (PE), sphingomyelin (SM), diacylglycerol (DG), phosphatidic acid (PA), and phosphatidylcholine (PC), which may provide insights into its pathogenesis and potential therapeutic targets. Furthermore, multi-omics network analysis revealed intricate metabolic pathways involved in GSRC progression. Our findings highlight the importance of understanding the metabolic heterogeneity of GSRC and pave the way for future studies exploring its clinical implications and therapeutic strategies.</p>","PeriodicalId":672,"journal":{"name":"Journal of the American Society for Mass Spectrometry","volume":" ","pages":""},"PeriodicalIF":3.1,"publicationDate":"2025-07-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144537692","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}

{"title":"Advanced Computational Approaches for Unraveling Transformation Pathways of Organic Micropollutants in Aquatic Environments: Insights from the Photodegradation of Ciprofloxacin.","authors":"Weiwen Sun, Xiaotong Geng, Jihong Xu, Yong Huang, Shifa Zhong, Lushi Lian, Xiaohong Guan","doi":"10.1021/jasms.5c00088","DOIUrl":"10.1021/jasms.5c00088","url":null,"abstract":"<p><p>Investigating the transformation products (TPs) of organic micropollutants in aquatic environments is crucial for understanding their fate, evaluating ecological and human health risks, and developing effective mitigation strategies to protect water quality. Thus, it is essential to establish an efficient workflow with low technical complexity for high-throughput TP identification. In this study, high resolution mass spectrometry (HRMS), stable isotope-labeled compounds, and similarity analysis were combined to develop an advanced computational approach for investigating the phototransformation processes of ciprofloxacin. A total of 68 tentatively ciprofloxacin-related TPs were extracted through isotope labeling experiments and formula filtering. Furthermore, structural elucidation of 42 TPs was achieved by combining HRMS/MS fragments and stable isotope labeling results, revealing that piperazinyl and cyclopropyl moieties are the key reaction sites of ciprofloxacin under solar irradiation. A novel similarity analysis workflow was developed to map the phototransformation pathways, establishing 80 parent-TP pairings. Dealkylation, oxygen addition, decarboxylation, and defluorination were found to be the dominant phototransformation reaction types of ciprofloxacin. The study highlights the complexity of ciprofloxacin phototransformation and provides a robust computational framework for elucidating the degradation pathways of organic contaminants. The developed methodology can be extended to study other emerging contaminants, supporting the design of more effective water treatment strategies and comprehensive risk assessments.</p>","PeriodicalId":672,"journal":{"name":"Journal of the American Society for Mass Spectrometry","volume":" ","pages":"1510-1517"},"PeriodicalIF":3.1,"publicationDate":"2025-07-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144324108","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}

{"title":"Energy-Dependent Changes in Ion Mobility Peak Width Reveal Stability Shifts in Ribonucleic Acid Stem Loops.","authors":"Anna G Anders, Jacqueline Anthenien, Ingrid R Kilde, Courteney Dufrene, Markos Koutmos, Brandon T Ruotolo","doi":"10.1021/jasms.5c00064","DOIUrl":"10.1021/jasms.5c00064","url":null,"abstract":"<p><p>Ribonucleic acids (RNAs) are becoming increasingly important to our understanding of cellular biology and our search for new or improved biotherapeutics. However, the dynamic and polydisperse nature of RNAs creates challenges for their structural characterization. Here, we focus on RNA stem-loops (i.e., hairpins), as these are ubiquitous elements of RNA secondary structures. We leverage native ion mobility-mass spectrometry (IM-MS) and collision-induced unfolding (CIU) to study stem loops isolated from mitochondrial tRNA (tRNA) across a wide range of disease-relevant mutants. While we find that native IM-MS analyses proved capable of tracking stem-loop structure changes upon cofactor-binding and mutation, it remains challenging to evaluate the stabilities of small RNAs using standard RNA CIU workflows. Thus, we explore the ability of energy-dependent changes by IM peak full-width half-maximum (fwhm) values to characterize the stabilities of RNA stem-loops. We find that RNA stem-loop IM fwhm values typically decrease significantly upon activation, with lesser differences detected in Mg²⁺-bound RNA samples in a manner consistent with the stabilization expected under such conditions. We conclude by discussing the potential future applications of IM fwhm analyses in the context of RNA CIU assay development.</p>","PeriodicalId":672,"journal":{"name":"Journal of the American Society for Mass Spectrometry","volume":" ","pages":"1422-1426"},"PeriodicalIF":3.1,"publicationDate":"2025-07-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144172136","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}

{"title":"Broadband Collision-Induced Dissociation Mass Spectrometry Imaging.","authors":"Sumi Krupa, Wiktoria Szuberla, Joanna Nizioł, Anna Ossolińska, Krzysztof Ossoliński, Tomasz Ruman","doi":"10.1021/jasms.5c00045","DOIUrl":"10.1021/jasms.5c00045","url":null,"abstract":"<p><p>This study demonstrates the first application of broadband collision-induced dissociation (bbCID) to mass spectrometry imaging (MSI) for the identification of untargeted metabolites in human tissues. The methodology integrates bbCID with laser ablation-remote atmospheric pressure photoionization/chemical ionization (LARAPPI/CI), enabling the simultaneous acquisition of precursor and fragment ion distributions during MSI for many compounds simultaneously. In this approach, an infrared (IR) laser is used to ablate biological material, which is then ionized in the gas phase by a combination of photoionization and chemical ionization at atmospheric pressure. The method was validated using reference compounds, including thymidine and commonly used synthetic dyes, to assess ionization efficiency, fragmentation behavior, and spatial colocalization of precursor and fragment ions. Subsequently, bbCID-MSI was applied to clinical tissue samples of human bladder and kidney cancer. For the bladder cancer tissue, higher intensities of heptadecanoic acid, docosahexaenoic acid (FA(22:6)), docosapentaenoic acid (FA(22:5)), and FA(16:0) were observed in tumor regions, whereas proline was more abundant in adjacent nontumorous area. In renal cell carcinoma, cancerous regions exhibited elevated levels of polyunsaturated fatty acids such as arachidonic acid (FA(20:4)) and adrenic acid (FA(22:4)), while creatine and serine were enriched in healthy tissue zones. These findings highlight the utility of bbCID-MSI for spatially resolved metabolite analysis and its potential to reveal biologically relevant metabolic alterations associated with cancer.</p>","PeriodicalId":672,"journal":{"name":"Journal of the American Society for Mass Spectrometry","volume":" ","pages":"1443-1455"},"PeriodicalIF":3.1,"publicationDate":"2025-07-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144324109","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}