Journal of the American Society for Mass Spectrometry最新文献

{"title":"Rapid Quantification of Neuraminidase Activity by MALDI-TOF MS via On-Target Labeling of Its Substrate and Product.","authors":"Jiarui Li, Xi Lin, Hao Wang, Nan Zhao, Xinhua Guo","doi":"10.1021/jasms.4c00446","DOIUrl":"https://doi.org/10.1021/jasms.4c00446","url":null,"abstract":"<p><p>Neuraminidase (NA) is a kind of glycoside hydrolase enzyme, functioning to remove terminal sialic acid (Sia) from glycans which are located on the cell surface. NA plays an essential role in cell interactions with ligands, microbes, and other cells during physiological and pathological processes. Additionally, NA is a major target for developing anti-influenza drugs and influenza vaccines. Herein, a matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) based method to quantify NA activity is demonstrated for the first time. A reactive matrix 2-hydrazinoquinoline (2-HQ) is used to on-target label the natural substrate (3-sialyllactose, 3'-SL) and its enzymatic product (Sia). The derivatization enhances the ionization efficiency of 3'-SL and Sia, especially in negative ion detection mode. Moreover, the lactose ion signals and noise are significantly suppressed. Consequently, NA activity in influenza vaccines has been successfully quantified by comparing the relative intensity of 2-HQ derivatized Sia and 3'-SL in the absence of an additional internal standard. Moreover, the inhibition efficiencies of NA inhibitors have also been measured. Due to its operating simplicity, high-throughput capacity, and quantification accuracy, the proposed method has potential to be applied for the detection of different kinds of NA activity to reveal the role of NA in immunity, vaccine, and infection processes.</p>","PeriodicalId":672,"journal":{"name":"Journal of the American Society for Mass Spectrometry","volume":" ","pages":""},"PeriodicalIF":3.1,"publicationDate":"2025-02-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143373718","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":"Use of TOFSim, a LabView-Based Time-of-Flight Mass Spectrometer Simulation, to Model Real Instrument Data.","authors":"Hannah M Palmer, Kevin G Owens","doi":"10.1021/jasms.4c00406","DOIUrl":"10.1021/jasms.4c00406","url":null,"abstract":"<p><p>It is demonstrated here that a recently published LabView-based time-of-flight mass spectrometer (TOFMS) simulation program (named TOFSim) can accurately simulate data collected on a commercial Bruker Autoflex III matrix-assisted laser desorption/ionization (MALDI) TOFMS instrument operating in linear mode. Once the instrument distances are determined by matching measured and simulated flight times, it is shown that both overall flight times and peak widths are reproduced for data collected under both focused and slightly defocused conditions. This work confirms that TOFSim can be used not just for training new instrument operators in the principles of TOFMS but, as demonstrated here, to show how changing the voltage applied to grid G1 in the source or the delayed extraction delay time affects the focusing properties of the instrument. In the future we expect that this will also allow users to perform \"what-if\" experiments to investigate scenarios which may be difficult or impossible to do in a real instrument.</p>","PeriodicalId":672,"journal":{"name":"Journal of the American Society for Mass Spectrometry","volume":" ","pages":""},"PeriodicalIF":3.1,"publicationDate":"2025-02-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143363518","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":"Rapid Removal of IgG1 Carryover on Protease Column Using Protease-Safe Wash Solutions Delivered with LC Pump for HDX-MS Systems.","authors":"Sandeep Kumar, Kyle W Anderson","doi":"10.1021/jasms.4c00411","DOIUrl":"10.1021/jasms.4c00411","url":null,"abstract":"<p><p>Sample carryover is a common problem in hydrogen-deuterium exchange mass spectrometry, particularly because immobilized protease columns cannot withstand the high organic solvent concentrations typically used in liquid chromatography-mass spectrometry (LC-MS) for cleaning. Conventional cleaning methods using injections of guanidine HCl still suffer from carryover and may require four blanks after each sample run to fully remove carryover. We have implemented an additional LC pump to deliver customized wash solutions to protease and enzyme columns, and the associated LC capillaries to eliminate carryover. Pump-based washing using the protease-safe wash solutions tested herein was able to fully remove carryover with only one blank run. FOS-choline-12 was found to be the most effective component in wash solutions and even performed well alone at 0.1% volume concentration. Since the protease column washing is performed concurrently during the analytical gradient within a sample run, subsequent blank runs could be reduced from four to one and total run time could be reduced by up to 60%. Savings in total run time could more than double the productivity of data acquisition, which is imperative for pandemic preparedness and for acceleration of biotherapeutics development.</p>","PeriodicalId":672,"journal":{"name":"Journal of the American Society for Mass Spectrometry","volume":" ","pages":"340-345"},"PeriodicalIF":3.1,"publicationDate":"2025-02-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142875527","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":"FAST MS: Software for the Automated Analysis of Top-Down Mass Spectra of Polymeric Molecules Including RNA, DNA, and Proteins.","authors":"Michael Palasser, Kathrin Breuker","doi":"10.1021/jasms.4c00236","DOIUrl":"10.1021/jasms.4c00236","url":null,"abstract":"<p><p>Top-down mass spectrometry (MS) enables comprehensive characterization of modified proteins and nucleic acids and, when native electrospray ionization (ESI) is used, binding site mapping of their complexes with native or therapeutic ligands. However, the high complexity of top-down MS spectra poses a serious challenge to both manual and automated data interpretation, even when the protein, RNA, or DNA sequence and the type of modification or the ligand are known. Here, we introduce FAST MS, a user-friendly software that identifies, assigns and relatively quantifies signals of molecular and fragment ions in MS and MS/MS spectra of biopolymers with known sequence and provides a toolbox for statistical analysis. FAST MS searches mass spectra for ion signals by comparing all signals in the spectrum with isotopic profiles calculated from known sequences, resulting in superior sensitivity and an increased number of assigned fragment ions compared to algorithms that rely on artificial monomer units while maintaining the false positive rate on a moderate level (<5%). FAST MS is an open-source, cross-platform software for the accurate identification, localization and relative quantification of modifications, even in complex mixtures of positional isomers of proteins, oligonucleotides, or any other user-defined linear polymer.</p>","PeriodicalId":672,"journal":{"name":"Journal of the American Society for Mass Spectrometry","volume":" ","pages":"247-257"},"PeriodicalIF":3.1,"publicationDate":"2025-02-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11808778/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142880971","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":"Characterization of Sugammadex-Related Isomeric Cyclodextrin Impurities Using Cyclic Ion Mobility High-Resolution Mass Spectrometry.","authors":"Péter S Szakály, Dávid Papp, Arnold Steckel, Erzsébet Varga, Gitta Schlosser","doi":"10.1021/jasms.4c00243","DOIUrl":"10.1021/jasms.4c00243","url":null,"abstract":"<p><p>Cyclic ion mobility-mass spectrometry (cIM-MS) is a powerful technique for separating and identifying isomeric mixtures of compounds. When coupled with chromatography, cIM-MS creates a multidimensional separation system, with high resolving power and peak capacity. In this study, we report the cyclic ion mobility separation and high-resolution mass spectrometry identification of four regioisomers of a Sugammadex-related impurity, abbreviated as Di-OH-SGM. Separation using multipass cyclic ion mobility was achieved by selecting the [M + 2Na]²⁺ ion, while other adducts, such as [M + Na]⁺, [M + 2H]²⁺, [M + H + Na]²⁺, and [M - 2H]^2- did not yield isomer separation. Two methods were developed for ion mobility separation of the isomers: a conventional multipass method and a slicing method. Isomer assignment was based on the characteristic fragment ions. The collision cross section values (^cTWCCS_N2) of the resolved cyclodextrin isomers were also determined. Ion mobility separation of structurally different fragment ions was demonstrated. Additionally, by coupling cIM-MS with reversed-phase liquid chromatography (HPLC-cIM-MS), two-dimensional separation of the isomers was achieved. The isomers, separated using HPLC-cIM-MS, were identified with the same approach as with cIM-MS alone, and their elution order provided insights into their relative hydrophobicity.</p>","PeriodicalId":672,"journal":{"name":"Journal of the American Society for Mass Spectrometry","volume":" ","pages":"258-264"},"PeriodicalIF":3.1,"publicationDate":"2025-02-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11808773/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143035827","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":"New Library-Based Methods for Nontargeted Compound Identification by GC-EI-MS.","authors":"Deborah F McGlynn, Lindsay D Yee, H Martin Garraffo, Lewis Y Geer, Tytus D Mak, Yuri A Mirokhin, Dmitrii V Tchekhovskoi, Coty N Jen, Allen H Goldstein, Anthony J Kearsley, Stephen E Stein","doi":"10.1021/jasms.4c00451","DOIUrl":"10.1021/jasms.4c00451","url":null,"abstract":"<p><p>While gas chromatography mass spectrometry (GC-MS) has long been used to identify compounds in complex mixtures, this process is often subjective and time-consuming and leaves a large fraction of seemingly good-quality spectra unidentified. In this work, we describe a set of new mass spectral library-based methods to assist compound identification in complex mixtures. These methods employ mass spectral uniqueness and compound ubiquity of library entries alongside noise reduction and automated comparison of retention indices to library compounds. As a test data set, we used a publicly available electron ionization mass spectrometry data set consisting of 4833 spectra of particulate organic compounds emitted by combustion of wildland fuels. In the present work, spectra in this data set were first identified using the NIST 2023 EI-MS Library and associated batch process identification software (NIST MS PepSearch) using retention-index corrected Identity Search scoring. Resulting identifications and related information were then employed to parametrize other factors that correlate with identification. A method for identifying compounds absent from but related to those present in mass spectral libraries using the Hybrid Similarity Search is illustrated. Nevertheless, some 90% of the spectra remain unidentified. Through comparison of unidentified to identified mass spectra in this data set, a new simple measure, namely median relative abundance, was developed for evaluating the likelihood of identification.</p>","PeriodicalId":672,"journal":{"name":"Journal of the American Society for Mass Spectrometry","volume":" ","pages":"389-399"},"PeriodicalIF":3.1,"publicationDate":"2025-02-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11844893/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142977063","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":"Ionization Characteristics of Glycan Homologues in Various Modes of Electrospray.","authors":"Nicholas R Allen, Kanwal Jeet, Tolulope Ogunsanya, Ian Ferraro, Nancy Fernandes, Huishan Li, Thaddaeus Webster, Carrie Mason, Anyin Li","doi":"10.1021/jasms.4c00425","DOIUrl":"10.1021/jasms.4c00425","url":null,"abstract":"<p><p>Fluorescence labeled glycan homologous mixtures were quantified using fluorescence and then used to evaluate ionization performances in electrospray ionization at micro, nano, and femto flow modes. nanoESI produced higher (2+ and 3+) charged ions adducted with sodium and calcium. In comparison, femtoESI was found to favor the generation of [M + H]⁺ ions against metal adducts, even with nonvolatile salts up to 1 mM for NaCl and 100 μM for CaCl₂. For labeled glucose homopolymer (GHP) glycans, nanoESI and femtoESI had 0.81 and 3 nM detection limits, respectively. With LC separation and a much higher flow rate, conventional microflow ESI detected all glycans with 10-fold lower concentrations. Overall, nanoESI had the optimum uniformity in the relative ionization efficiency (RIE). When summing up intensities of analyte ions formed with all charge carriers, the RIE of the midsized glycans (10 to 16 glucose units) appear to be uniform (RIE 95%-105%). For the smaller (1-5 glucose units) glycan components, femtoESI provided better uniformity than nanoESI and conventional ESI. For the labeled IgG N-glycans, the impact of chemical structure on the ionization efficiency was revealed by the strong correlation between their RIE trends in different ionization modes.</p>","PeriodicalId":672,"journal":{"name":"Journal of the American Society for Mass Spectrometry","volume":" ","pages":"346-354"},"PeriodicalIF":3.1,"publicationDate":"2025-02-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142875517","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":"Gas-Phase Fluorescence Excitation Experiments on Cryogenically Cold Rhodamine B Cations Linked to Various Amino Acid Esters.","authors":"Iden Djavani-Tabrizi, Ziqi Yuan, Thomas Toft Lindkvist, Jiamin Xing, Li Chen, Steen Bro Ndsted Nielsen","doi":"10.1021/jasms.4c00469","DOIUrl":"10.1021/jasms.4c00469","url":null,"abstract":"<p><p>Förster resonance energy transfer (FRET) is becoming a valuable technique in gas-phase structural biology for identifying local structural motifs and conformations of biological molecules, such as peptides and proteins. This method involves labeling the biomolecule with two dyes, a donor dye and an acceptor dye, that are commonly charged rhodamines. Here we examine how different amino acid (AA) methyl esters linked to the dye via amide linkages can influence the dye transition energy and, consequently, the energy-transfer efficiency, using cryogenic ion fluorescence spectroscopy. Absorption spectra were recorded for rhodamine B⁺-labeled AA esters (RB⁺-AA) through fluorescence-excitation experiments at the LUNA2 setup in Aarhus, which operates at cryogenic temperatures (down to approximately 100 K). The AAs studied include aliphatic ones (alanine (A), leucine (L), <i>tert</i>-leucine (<i>tert</i>-L), and methionine (M)), aromatic ones (phenylalanine (F) and tryptophan (W)), and two with polar side chains (serine (S) and threonine (T)). Results show that the band maximum either remains unchanged compared to RB⁺ or red shifts by over 3 nm in the case of RB⁺-M and RB⁺-F. While the spectra of RB⁺-A and RB⁺-L closely resemble that of RB⁺, RB⁺-<i>tert</i>-L shows a distinct red shift of about 1.4 nm. Spectral variations do not appear to be more influenced by the presence of aromatic AA side chains than other types, as differences observed between aliphatic AAs are comparable to those between the three groups. Instead, these variations appear to arise from differing conformations where the dihedral angle between the xanthene moiety and the pendant phenyl group varies, as influenced by the linked AA side chain. The angle determines the π-overlap between the two aromatic moieties, and according to TD-DFT calculations, an angle larger than 90° can easily account for red shifts due to larger delocalization of the π-electron cloud. Another factor is the polarizability of the side chain that could also contribute to the red shift. RB⁺-F and RB⁺-W spectra exhibit red-shifted, narrower absorption profiles, which is likely associated with the large aromatic side chains that limit the number of contributing structural configurations.</p>","PeriodicalId":672,"journal":{"name":"Journal of the American Society for Mass Spectrometry","volume":" ","pages":"409-415"},"PeriodicalIF":3.1,"publicationDate":"2025-02-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142930295","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":"Mass Spectrometry Analysis of Chemically and Collisionally Dissociated Molecular Glue- and PROTAC-Mediated Protein Complexes Informs on Disassembly Pathways.","authors":"Edvaldo V S Maciel, Jonathan Eisert, Julian Müller, Tanja Habeck, Frederik Lermyte","doi":"10.1021/jasms.4c00429","DOIUrl":"10.1021/jasms.4c00429","url":null,"abstract":"<p><p>Molecular glues (MGs) and proteolysis-targeting chimeras (PROTACs) are used to modulate protein-protein interactions (PPIs), via induced proximity between compounds that have little or no affinity for each other naturally. They promote either reversible inhibition or selective degradation of a target protein, including ones deemed undruggable by traditional therapeutics. Though native MS (nMS) is capable of analyzing multiprotein complexes, the behavior of these artificially induced compounds in the gas phase is still not fully understood, and the number of publications over the past few years is still rather limited. Here, we studied two MG-induced complexes between mTOR_FRB and FKBP12 as well as a PROTAC-induced complex between FKBP51_FK1 and the von Hippel-Lindau E3 ligase (VHL). Native MS combined with collision-induced dissociation (CID) provided a way of measuring not only the formation of these complexes but also their dissociation pathways. Both protein complexes seem to eject preferably the centrally located small (compared to the mass of the proteins) ligand upon CID, rather than dissociating a peripheral subunit, as is often observed for naturally occurring protein complexes. In contrast, chemically induced dissociation in solution generated complementary data to CID, by disrupting the PPI surface, which resulted in more diverse MS spectra that preserved the stronger interactions in solution.</p>","PeriodicalId":672,"journal":{"name":"Journal of the American Society for Mass Spectrometry","volume":" ","pages":"355-367"},"PeriodicalIF":3.1,"publicationDate":"2025-02-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142982469","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":"Rigorous Analysis of Multimodal HDX-MS Spectra.","authors":"Lisa M Tuttle, Ellie I James, Florian Georgescauld, Thomas E Wales, David D Weis, John R Engen, Abhinav Nath, Rachel E Klevit, Miklos Guttman","doi":"10.1021/jasms.4c00471","DOIUrl":"10.1021/jasms.4c00471","url":null,"abstract":"<p><p>An inherent strength of hydrogen/deuterium exchange coupled to mass spectrometry (HDX-MS) is its ability to detect the presence of multiple conformational states of a protein, which often manifest as multimodal isotopic envelopes. However, the statistical considerations for accurate analysis of multimodal spectra have yet to be established. Here we outline an unrestrained binomial distribution fitting approach with the corresponding statistical tests to accurately detect and, when possible, deconvolute isotopic distributions that contain multiple subpopulations. The algorithms have been incorporated into an updated version of the freely available software, HX-Express, and validated using known mixtures of peptides deuterated to varying degrees. This approach presents a readily accessible tool to fit and interpret bimodal and trimodal behavior in HDX-MS data for mixed populations, EX1 kinetics, and pulse labeling data.</p>","PeriodicalId":672,"journal":{"name":"Journal of the American Society for Mass Spectrometry","volume":" ","pages":"416-423"},"PeriodicalIF":3.1,"publicationDate":"2025-02-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142998195","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}