Journal of the American Society for Mass Spectrometry最新文献

{"title":"Mapping Binding Domains of Viral and Allergenic Proteins with Dual-Cleavable Cross-Linking Technology.","authors":"Akash Talukder, Saiful M Chowdhury","doi":"10.1021/jasms.4c00398","DOIUrl":"10.1021/jasms.4c00398","url":null,"abstract":"<p><p>The dual-cleavable nature of the cross-linking technology (DUCCT) enhances the reliable identification of cross-linked peptides via mass spectrometry. The DUCCT approach uses a cross-linking agent that can be selectively cleaved by two different tandem mass spectrometry techniques: collision-induced dissociation (CID) and electron transfer dissociation (ETD). This results in distinct signatures in two independent mass spectra for the same cross-linked precursor, leading to unambiguous identification and the validation of the spectra. In this study, we expanded the application of the DUCCT cross-linker to evaluate the binding domains of a specific cat dander allergen, Fel d 1, which exists as the Fel d 1 A and B protein complex, and a viral spike protein from SARS-CoV-2, which invades host cells. To assess the cross-linked products obtained by DUCCT, we utilized a software tool called Cleave-XL, which effectively identified cross-linked sites using data from CID and ETD. Dual cleavable cross-linking studies identified cross-linked peptides in these complexes, which have been reported in bioinformatics analysis and proposed for immunotherapy using synthetic peptides. A benchmark study was also conducted using a commercial cross-linker disuccinimidyl suberate (DSS). Overall, we expect that DUCCT cross-linking technology will greatly facilitate the rapid screening of binding interfaces, thereby advancing structural biology and cell signaling investigations.</p>","PeriodicalId":672,"journal":{"name":"Journal of the American Society for Mass Spectrometry","volume":" ","pages":"721-731"},"PeriodicalIF":3.1,"publicationDate":"2025-04-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143690766","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":"Bromine-Based Derivatization of Carboxyl-Containing Metabolites for Liquid Chromatography-Trapped Ion Mobility Spectrometry-Mass Spectrometry.","authors":"Kaylie I Kirkwood-Donelson, Prashant Rai, Lalith Perera, Michael B Fessler, Alan K Jarmusch","doi":"10.1021/jasms.5c00023","DOIUrl":"10.1021/jasms.5c00023","url":null,"abstract":"<p><p>The analysis of small carboxyl-containing metabolites (CCMs), such as tricarboxylic acid (TCA) cycle intermediates, provides highly useful information about the metabolic state of cells. However, their detection using liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS) methods can face sensitivity and specificity challenges given their low ionization efficiency and the presence of isomers. Ion mobility spectrometry (IMS), such as trapped ion mobility spectrometry (TIMS), provides additional specificity, but further signal loss can occur during the mobility separation process. We, therefore, developed a solution to boost CCM ionization and chromatographic separation as well as leverage specificity of IMS. Inspired by carbodiimide-mediated coupling of carboxylic acids with 4-bromo-<i>N</i>-methylbenzylamine (4-BNMA) for quantitative analysis, we newly report the benefits of this reagent for TIMS-based measurement. We observed a pronounced (orders of magnitude) increase in signal and enhanced isomer separations, particularly by LC. We found that utilization of a brominated reagent, such as 4-BNMA, offered unique benefits for untargeted CCM measurement. Derivatized CCMs displayed shifted mobility out of the metabolite and lipid region of the TIMS-MS space as well as characteristic isotope patterns, which were leveraged for data mining with Mass Spectrometry Query Language (MassQL) and indication of the number of carboxyl groups. The utility of our LC-ESI-TIMS-MS/MS method with 4-BMA derivatization was demonstrated via the characterization of alterations in CCM expression in bone marrow-derived macrophages upon activation with lipopolysaccharide. While metabolic reprogramming in activated macrophages has been characterized previously, especially with respect to TCA cycle intermediates, we report a novel finding that isomeric itaconic, mesaconic, and citraconic acid increase after 24 h, indicating possible roles in the inflammatory response.</p>","PeriodicalId":672,"journal":{"name":"Journal of the American Society for Mass Spectrometry","volume":" ","pages":"888-899"},"PeriodicalIF":3.1,"publicationDate":"2025-04-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11970421/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143571911","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":"Spatial Distribution of Brain PET Tracers by MALDI Imaging.","authors":"Isabeau Vermeulen, Michiel Vandenbosch, Delphine Viot, Joel Mercier, Diego Asensio-Wandosell Cabañas, Pilar Martinez-Martinez, Patrick Barton, Ron M A Heeren, Berta Cillero-Pastor","doi":"10.1021/jasms.4c00307","DOIUrl":"10.1021/jasms.4c00307","url":null,"abstract":"<p><p>Evaluating tissue distribution of Positron Emission Tomography (PET) tracers during their development conventionally involves autoradiography techniques, where radioactive compounds are used for <i>ex vivo</i> visualization and quantification in tissues during preclinical development stages. Mass Spectrometry Imaging (MSI) offers a potential alternative, providing spatial information without the need for radioactivity with a similar spatial resolution. This study aimed to optimize a MSI sample preparation protocol for assessing PET tracer candidates <i>ex vivo</i> with a focus on two compounds: UCB-J and UCB2400. We tested different matrices and introduced washing steps to improve PET tracer detection. Tissue homogenates were prepared to construct calibration curves for quantification. The incorporation of a washing step into the MSI sample preparation protocol enhanced the signal of both PET tracers. Our findings highlight MSI's potential as a cost-effective and efficient method for the evaluation of PET tracer distribution. The optimized approach offered here can provide a protocol that enhances the signal and minimizes ion suppression effect, which can be valuable for future evaluation of PET tracers in MSI studies.</p>","PeriodicalId":672,"journal":{"name":"Journal of the American Society for Mass Spectrometry","volume":" ","pages":"688-698"},"PeriodicalIF":3.1,"publicationDate":"2025-04-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11969657/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143612917","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":"Glycosaminoglycan Mass Spectrometry Imaging by Infrared Matrix-Assisted Laser Desorption Electrospray Ionization.","authors":"Tana V Palomino, David C Muddiman","doi":"10.1021/jasms.4c00435","DOIUrl":"10.1021/jasms.4c00435","url":null,"abstract":"<p><p>Chondroitin sulfate (CS) is a type of glycosaminoglycan (GAG) that is abundant in cartilage and perineural networks (PNNs). Changes in the CS signature of PNNs have been implicated in several neurological diseases. Most CS-GAGs contain labile sulfate groups, which can be lost during ionization events that deposit large amounts of internal energy. Infrared matrix-assisted laser desorption electrospray ionization (IR-MALDESI) is a soft ionization technique used for mass spectrometry imaging. In this work, we determine the spatial distribution of CS-GAG disaccharides within rodent brain using IR-MALDESI MSI. Non-, mono-, and disulfated disaccharides were detected with various adducts. All disaccharides colocalized to the PNNs, which are most abundant in the cortex and hippocampus regions of the brain. This is the first MSI study to spatially resolve CS-GAG disaccharides within brain, paving the way for IR-MALDESI to measure GAGs in neurological diseases.</p>","PeriodicalId":672,"journal":{"name":"Journal of the American Society for Mass Spectrometry","volume":" ","pages":"658-663"},"PeriodicalIF":3.1,"publicationDate":"2025-04-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143539917","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":"Detection and Structural Elucidation of Copper Binding Tri- and Tetrapyrrole Ligands Produced by the Marine Diatom <i>Phaeodactylum Tricornutum</i>.","authors":"Lydia Babcock-Adams, Jingxuan Li, Amy M McKenna, Christopher L Hendrickson, Daniel J Repeta","doi":"10.1021/jasms.4c00380","DOIUrl":"10.1021/jasms.4c00380","url":null,"abstract":"<p><p>In seawater, most dissolved copper (Cu) is complexed by organic ligands, many of which are thought to be produced by phytoplankton. Although very little is known about the composition and structure of these ligands, they play an important role in determining the reactivity and bioavailability of Cu. In this study, <i>Phaeodactylum tricornutum</i>, a marine diatom known to produce Cu ligands (CuLs), was grown in laboratory pure culture, and the CuLs were recovered from the growth media. Using liquid chromatography coupled to ultrahigh resolution tandem mass spectrometry, 11 Cu ligand complexes were identified and assigned molecular formulas. Molecular formulas were confirmed by comparing the expected and observed relative abundances of ¹⁵N, ¹³C, ⁶⁵Cu, and ¹⁸O isotopologues. The CuLs had molecular weights from 520 to 719 Da and molecular formulas of C_26-35H_23-36O_5-9N_3-4Cu with an average assignment error of 56 ppb. High-resolution tandem mass spectrometry of the Cu-bound and metal-free ligands revealed these to be a suite of tri- and tetrapyrroles stabilized through complexation of Cu by N. The ligands share similar parent structures but differ in the number, type, and arrangement of functional groups that decorate the pyrroles. The similarity of CuL structures with known catabolites of chlorophyll suggests these ligands may be widely produced by marine photoautotrophs.</p>","PeriodicalId":672,"journal":{"name":"Journal of the American Society for Mass Spectrometry","volume":" ","pages":"713-720"},"PeriodicalIF":3.1,"publicationDate":"2025-04-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11970420/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143405088","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":"Pneumatically Assisted Microfluidic Probe for Enhanced Mass Spectrometry Imaging Performance.","authors":"Li-Xue Jiang, Julia Laskin","doi":"10.1021/jasms.5c00011","DOIUrl":"10.1021/jasms.5c00011","url":null,"abstract":"<p><p>A pneumatically assisted microfluidic probe (MFP) with two microfluidic channels has been developed for nanospray desorption electrospray ionization mass spectrometry imaging (nano-DESI MSI) of biological samples. This design simplifies the experimental setup, making it independent of the vacuum suction at the mass spectrometer inlet. The implementation of pneumatically assisted solvent flow through the probe enables stable, high solvent flow rates required to maintain a consistent liquid bridge during high-throughput MSI experiments. This approach addresses challenges associated with using MFP nano-DESI probes on mass spectrometers that have limited vacuum suction and the operation of MFPs with small microfluidic channels. We demonstrate the robustness of the pneumatically assisted MFP with 30 μm channels, which cannot be used for high-throughput MSI experiments without pneumatic assistance, by successfully imaging five mouse brain tissue sections without interruptions.</p>","PeriodicalId":672,"journal":{"name":"Journal of the American Society for Mass Spectrometry","volume":" ","pages":"883-887"},"PeriodicalIF":3.1,"publicationDate":"2025-04-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143668541","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":"Structural Characterization of Dimeric Perfluoroalkyl Carboxylic Acid Using Experimental and Theoretical Ion Mobility Spectrometry Analyses.","authors":"Aurore L Schneiders, Johann Far, Lidia Belova, Allison Fry, Adrian Covaci, Erin S Baker, Edwin De Pauw, Gauthier Eppe","doi":"10.1021/jasms.5c00007","DOIUrl":"10.1021/jasms.5c00007","url":null,"abstract":"<p><p>Per- and polyfluoroalkyl substances (PFAS) are contaminants of increasing concern, with over seven million compounds currently inventoried in the PubChem PFAS Tree. Recently, ion mobility spectrometry has been combined with liquid chromatography and high-resolution mass spectrometry (LC-IMS-HRMS) to assess PFAS. Interestingly, using negative electrospray ionization, perfluoroalkyl carboxylic acids (PFCAs) form homodimers ([2M-H]^-), a phenomenon observed with trapped, traveling wave, and drift-tube IMS. In addition to the limited research on their effect on analytical performance, there is little information on the conformations these dimers can adopt. This study aimed to propose most probable conformations for PFCA dimers. Based on qualitative analysis of how collision cross section (CCS) values change with the mass-to-charge ratio (<i>m</i>/<i>z</i>) of PFCA ions, the PFCA dimers were hypothesized to likely adopt a V-shaped structure. To support this assumption, <i>in silico</i> geometry optimizations were performed to generate a set of conformers for each possible dimer. A CCS value was then calculated for each conformer using the trajectory method with Lennard-Jones and ion-quadrupole potentials. Among these conformers, at least one of the ten lowest-energy conformers identified for each dimer exhibited theoretical CCS values within a ±2% error margin compared to the experimental data, qualifying them as plausible structures for the dimers. Our findings revealed that the fluorinated alkyl chains in the dimers are close to each other due to a combination of C-F···O=C and C-F···F-C stabilizing interactions. These findings, together with supplementary investigations involving environmentally relevant cations, may offer valuable insights into the interactions and environmental behavior of PFAS.</p>","PeriodicalId":672,"journal":{"name":"Journal of the American Society for Mass Spectrometry","volume":" ","pages":"850-861"},"PeriodicalIF":3.1,"publicationDate":"2025-04-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143565636","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":"Faces of Mass Spectrometry/Amanda Patrick.","authors":"Anne Brenner, J D Brookbank","doi":"10.1021/jasms.5c00042","DOIUrl":"10.1021/jasms.5c00042","url":null,"abstract":"","PeriodicalId":672,"journal":{"name":"Journal of the American Society for Mass Spectrometry","volume":" ","pages":"628-631"},"PeriodicalIF":3.1,"publicationDate":"2025-04-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143490260","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":"Headspace Injection Method for Intermittent Sampling and Profiling Analyses of Volatile Organic Compounds Using Dielectric Barrier Discharge Ionization (DBDI).","authors":"Daniel Heffernan, Frederik Oleinek, Ayla Schueler, Paak Wai Lau, Jürgen Kudermann, Alina Meindl, Mathias O Senge, Nicole Strittmatter","doi":"10.1021/jasms.4c00475","DOIUrl":"10.1021/jasms.4c00475","url":null,"abstract":"<p><p>A direct headspace injection method is presented and optimized for the analysis of volatile organic compounds (VOCs) using dielectric barrier discharge ionization-mass spectrometry (DBDI-MS), incorporating an intermediate vial in which the sample headspace is injected. The setup is built of commonly available, cheap consumable parts and easily enables the incorporation of different gases for generating different ionization atmospheres. The method can be fully automated by using standard GC autosamplers, and its rapid analysis time is suitable for high-throughput applications. We show that this method is suitable for both profiling analysis of complex samples such as biofluids and quantitative measurements for real-time reaction monitoring. Our optimized method demonstrated improved reproducibility and sensitivity, with detection limits for compounds tested in the high nanomolar to the low micromolar range, depending on the compound. Key parameters for method optimization were identified such as sample vial volume, headspace-to-liquid ratio, incubation temperature, and equilibration time. These settings were systematically evaluated to maximize the signal intensity and improve repeatability between measurements. Two use cases are demonstrated: (i) quantitative measurement of ethanol production by a metal-organic framework from CO₂ and (ii) profiling of biofluids following the consumption of asparagus.</p>","PeriodicalId":672,"journal":{"name":"Journal of the American Society for Mass Spectrometry","volume":" ","pages":"801-810"},"PeriodicalIF":3.1,"publicationDate":"2025-04-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11969650/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143596010","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":"Kinetic Method Coupled with Thermal-Assisted Paper Spray Ionization Mass Spectrometry for Direct Determination of Enantiomeric Excess of Multiple d/l-Amino Acids in Functional Foods.","authors":"Wen-Bo Gao, Shu-Ting Xu, Yong-Jie Yan, Cheng Yang, Xiu-Ping Yan","doi":"10.1021/jasms.5c00050","DOIUrl":"10.1021/jasms.5c00050","url":null,"abstract":"<p><p>Amino acids are commonly used as nutritional fortification substances in functional foods, and their chiral configuration is an important determinant of food function. Rapid chiral screening methods are urgently needed in food analysis but are limited by the long-time chiral separation and matrix interference. In this study, we show a kinetic method coupled to thermal-assisted paper spray ionization mass spectrometry for direct determination of enantiomeric excess (<i>ee</i>) of multiple d/l-amino acids in complex food matrixes without sample pretreatment. 3-(2-Naphthyl)-l-alanine was selected as a new chiral reference ligand for the kinetic method to achieve efficient chiral differentiation (discrimination degree is 8.7 for d/l-phenylalanine and 10.2 for d/l-tyrosine). An additional thermal-auxiliary device was developed for paper spray ionization mass spectrometry to facilitate the enantiomeric purity determination. The developed method allowed a rapid simultaneous enantiomeric purity determination of multiple chiral amino acids (d/l-phenylalanine and d/l-tyrosine) within 30 s. Good linearities were achieved for the quantitation of <i>ee</i> (<i>R</i>² = 0.9996 for phenylalanine and 0.9995 for tyrosine) with unknown amino acid concentrations ranging from 10 μM to 600 μM. The developed method was successfully applied for the enantiomeric purity determination of multiple chiral amino acids in functional capsules and beverages and showed great potential for efficient enantiomer-related food safety screening and nutrition analysis.</p>","PeriodicalId":672,"journal":{"name":"Journal of the American Society for Mass Spectrometry","volume":" ","pages":"906-913"},"PeriodicalIF":3.1,"publicationDate":"2025-04-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143622920","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}