Linkage Memory in Underivatized Protonated Carbohydrates

Mookherjee, Abhigya; Uppal, Sanjit S.; Murphree, Taylor A.; Guttman, Miklós

doi:10.1021/jasms.0c00440

Cited by 11 publications

(14 citation statements)

References 51 publications

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“…Previous literature reports have shown that Fuc(α1,2)Gal(β1,4)Glc and Gal(β1,4)-[Fuc(α1,3)]-Glc could not be resolved from one another with a homebuilt 2 m DT IMS–MS platform but did have diagnostic cryogenic IR fingerprints. Another recent study on the three fucosylglucosamine isomers showed that subtle differences were present in their arrival time distributions as their [M – H 2 O + H] + adducts on a 25 cm TWIMS–MS platform but were not analyzed, or resolved, as an isomeric mixture. By utilizing the novel capabilities of the cIMS array, we were able to slice out selected mobility regions and subject them to additional cycles, thus enabling the resolution of these challenging fucose-containing HMO building block isomers.…”

Section: Results and Discussionmentioning

confidence: 99%

Toward Sequencing the Human Milk Glycome: High-Resolution Cyclic Ion Mobility Separations of Core Human Milk Oligosaccharide Building Blocks

Peterson

Nagy

2021

Anal. Chem.

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Human milk oligosaccharides (HMOs) are an unconjugated class of glycans that have been implicated for their role in promoting the healthy development of the brain–gut axes of infants. Production of HMOs is ever-changing and specifically tailored for each infant in response to various biological factors (e.g., cognitive development, diseases, or allergies). While every HMO consists of up to only five monosaccharides, their structures can be composed of many possible glycosidic linkage positions and corresponding α/β anomericities, linear or branched chains, and potential fucosylation/sialylation modifications, thus leading to a tremendous degree of isomeric heterogeneity. With limited availability of authentic standards for every putative HMO structure (estimated to be >200 total), new analytical methods are needed for their accurate characterization. Complete sequencing of the human milk glycome would enable a better understanding of their infant-specific biological roles and potentially lead to their widespread incorporation into infant formula. Herein, we explore the use of our high-resolution cyclic ion mobility spectrometry–mass spectrometry (cIMS–MS)-based platform for the separation of core disaccharide and trisaccharide isomer building blocks as a first step toward the sequencing of larger HMOs. By utilizing the flexible capabilities of the cIMS array, separation pathlengths were extended up to 40 m, thus enabling the resolution of all seven sets of sialylated, fucosylated galactosyllactose and lactosamine HMO building block isomers. Additionally, we assessed the utility of pre-/post-cIMS tandem mass spectrometry (MS/MS) and tandem cIMS (cIMS/cIMS) for the characterization of HMOs based on their diagnostic fragmentation patterns and mobility fingerprints. We anticipate that our presented cIMS–MS-based methodology will enable the better characterization of larger, unknown HMOs when incorporated into an overall workflow that also includes online liquid chromatography and enzymatic hydrolyses.

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Section: Results and Discussionmentioning

confidence: 99%

Toward Sequencing the Human Milk Glycome: High-Resolution Cyclic Ion Mobility Separations of Core Human Milk Oligosaccharide Building Blocks

Peterson

Nagy

2021

Anal. Chem.

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“…Glycosidic cleavage yields information about glycan sequence, whereas cross-ring cleavage is important to deduce information about linkage and branching. Glycosidic B-fragments are also particularly interesting for chemical synthesis of glycans as they are believed to occur as intermediates of S N 1 reactions in solution, and their reactivity can be predicted by gas-phase studies. , Glycosidic B- and C-fragments showed in some cases a memory of the stereoinformation on the glycosidic linkage in unprotected glycosides − but not in protected glycosides …”

Section: Techniquesmentioning

confidence: 99%

Mass Spectrometry-Based Techniques to Elucidate the Sugar Code

et al. 2021

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Cells encode information in the sequence of biopolymers, such as nucleic acids, proteins, and glycans. Although glycans are essential to all living organisms, surprisingly little is known about the “sugar code” and the biological roles of these molecules. The reason glycobiology lags behind its counterparts dealing with nucleic acids and proteins lies in the complexity of carbohydrate structures, which renders their analysis extremely challenging. Building blocks that may differ only in the configuration of a single stereocenter, combined with the vast possibilities to connect monosaccharide units, lead to an immense variety of isomers, which poses a formidable challenge to conventional mass spectrometry. In recent years, however, a combination of innovative ion activation methods, commercialization of ion mobility–mass spectrometry, progress in gas-phase ion spectroscopy, and advances in computational chemistry have led to a revolution in mass spectrometry-based glycan analysis. The present review focuses on the above techniques that expanded the traditional glycomics toolkit and provided spectacular insight into the structure of these fascinating biomolecules. To emphasize the specific challenges associated with them, major classes of mammalian glycans are discussed in separate sections. By doing so, we aim to put the spotlight on the most important element of glycobiology: the glycans themselves.

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“…Ion mobility spectrometry (IMS), a technique in which gas-phase ions are separated based on their collisional cross section with an inert buffer gas, has been combined with MS n to provide a rapid and effective tool for glycan analysis. − Such studies in which fragments generated by collision-induced dissociation (CID) are characterized by IMS have led to the identification of regioisomers and anomers of the precursor glycans. , Ultrahigh-resolution IMS has been able to separate subtle substructures in small glycans, − and a cyclic IMS instrument has recently been used to separate and partially identify the isomers of a set of pentasaccharides based on their fragmentation pattern. , However, the ever-increasing resolution of IMS devices leads to the observation of multiple peaks in the arrival-time distribution (ATD). This complicates the interpretation of IMS data, as the separated reducing-end anomers, which are in equilibrium in solution, can then be confused with other structures such as positional isomers, regioisomers, or conformers.…”

Section: Introductionmentioning

confidence: 99%

A New Strategy Coupling Ion-Mobility-Selective CID and Cryogenic IR Spectroscopy to Identify Glycan Anomers

Pellegrinelli

Yue

Carrascosa

et al. 2022

J. Am. Soc. Mass Spectrom.

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Determining the primary structure of glycans remains challenging due to their isomeric complexity. While high-resolution ion mobility spectrometry (IMS) has recently allowed distinguishing between many glycan isomers, the arrival-time distributions (ATDs) frequently exhibit multiple peaks, which can arise from positional isomers, reducing-end anomers, or different conformations. Here, we present the combination of ultrahigh-resolution ion mobility, collision-induced dissociation (CID), and cryogenic infrared (IR) spectroscopy as a systematic method to identify reducing-end anomers of glycans. Previous studies have suggested that high-resolution ion mobility of sodiated glycans is able to separate the two reducing-end anomers. In this case, Y-fragments generated from mobility-separated precursor species should also contain a single anomer at their reducing end. We confirm that this is the case by comparing the IR spectra of selected Y-fragments to those of anomerically pure mono- and disaccharides, allowing the assignment of the mobility-separated precursor and its IR spectrum to a single reducing-end anomer. The anomerically pure precursor glycans can henceforth be rapidly identified on the basis of their IR spectrum alone, allowing them to be distinguished from other isomeric forms.

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Linkage Memory in Underivatized Protonated Carbohydrates

Cited by 11 publications

References 51 publications

Toward Sequencing the Human Milk Glycome: High-Resolution Cyclic Ion Mobility Separations of Core Human Milk Oligosaccharide Building Blocks

Toward Sequencing the Human Milk Glycome: High-Resolution Cyclic Ion Mobility Separations of Core Human Milk Oligosaccharide Building Blocks

Mass Spectrometry-Based Techniques to Elucidate the Sugar Code

A New Strategy Coupling Ion-Mobility-Selective CID and Cryogenic IR Spectroscopy to Identify Glycan Anomers

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