Acid‐catalyzed reductive amination of aldoses with 8‐aminopyrene‐1,3,6‐trisulfonate

Evangelista, Ramon A.; Guttman, András; Chen, Fu Tai A.

doi:10.1002/elps.1150170210

Cited by 57 publications

(35 citation statements)

References 18 publications

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“…Organic acids with low pK a , such as acetic acid (pK a = 4.75), malonic acid (pK a1 = 2.83) or citric acid (pK a1 = 3.15), are more commonly used to accelerate glycan labeling [27,28]. Studies have shown that stronger acids were able to increase derivatization yield [29], but their use was associated with higher sialic acid loss [30]. Among the numerous sugar labeling dyes the 8-aminopyrene-1,3,6-trisulfonic acid (APTS) [7] and 8-aminonaphthalene-1,3,6-trisulfonic (ANTS) [31] are the mostly used fluorophores in sugar analysis using electric field mediated separation methods as they are multiply charged and provide high fluorescent yield [32].…”

Section: Introductionmentioning

confidence: 99%

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Evaporative fluorophore labeling of carbohydrates via reductive amination

Reider

Szigeti

Guttman

2018

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As analytical glycomics became to prominence, newer and more efficient sample preparation methods are being developed. Albeit, numerous reductive amination based carbohydrate labeling protocols have been reported in the literature, the preferred way to conduct the reaction is in closed vials. Here we report on a novel evaporative labeling protocol with the great advantage of continuously concentrating the reagents during the tagging reaction, therefore accommodating to reach the optimal reagent concentrations for a wide range of glycan structures in a complex mixture. The optimized conditions of the evaporative labeling process minimized sialylation loss, otherwise representing a major issue in reductive amination based carbohydrate tagging. In addition, complete and uniform dispersion of dry samples was obtained by supplementing the low volume labeling mixtures (several microliters) with the addition of extra solvent (e.g., THF).Evaporative labeling is an automation-friendly glycan labeling method, suitable for standard open 96 well plate format operation. Graphical abstract 2/13

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Section: Introductionmentioning

confidence: 99%

“…Procedures reported in the literature apply comparable but not unified reaction conditions and reagent volumes for fluorescent sugar labeling [10], most of them suggesting overnight incubation at 37°C [29,33] or several hours of reaction times at higher temperatures (50°C) [34].…”

Section: Introductionmentioning

confidence: 99%

Evaporative fluorophore labeling of carbohydrates via reductive amination

Reider

Szigeti

Guttman

2018

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“…Lyophilized oligosaccharide standards, Man-9, Man-5, Man-3a, and Man-1 were labeled with APTS by reductive amination exactly as previously described (8). Excess labeling reagents and impurities were removed from APTS-labeled standards by fluorophore-assisted carbohydrate electrophoresis as described previously (9) with minor modifications (supplemental Fig.…”

Section: Methodsmentioning

confidence: 99%

Analysis of a New Family of Widely Distributed Metal-independent α-Mannosidases Provides Unique Insight into the Processing of N-Linked Glycans

Gregg

Zandberg

Hehemann

et al. 2011

Journal of Biological Chemistry

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The modification of N-glycans by ␣-mannosidases is a process that is relevant to a large number of biologically important processes, including infection by microbial pathogens and colonization by microbial symbionts. At present, the described mannosidases specific for ␣1,6-mannose linkages are very limited in number. Through structural and functional analysis of two sequence-related enzymes, one from Streptococcus pneumoniae (SpGH125) and one from Clostridium perfringens (CpGH125), a new glycoside hydrolase family, GH125, is identified and characterized. Analysis of SpGH125 and CpGH125 reveal them to have exo-␣1,6-mannosidase activity consistent with specificity for N-linked glycans having their ␣1,3-mannose branches removed. The x-ray crystal structures of SpGH125 and CpGH125 obtained in apo-, inhibitor-bound, and substratebound forms provide both mechanistic and molecular insight into how these proteins, which adopt an (␣/␣) 6 -fold, recognize and hydrolyze the ␣1,6-mannosidic bond by an inverting, metal-independent catalytic mechanism. A phylogenetic analysis of GH125 proteins reveals this to be a relatively large and widespread family found frequently in bacterial pathogens, bacterial human gut symbionts, and a variety of fungi. Based on these studies we predict this family of enzymes will primarily comprise such exo-␣1,6-mannosidases.A feature of emerging importance to bacteria that colonize or infect humans is their capacity to process host glycans. Streptococcus pneumoniae is one notable human pathogen that relies on this ability for its full virulence (1). Among its known carbohydrate active virulence factors are NanA, StrH, BgaA, and EndoD. NanA Glycoside hydrolases, enzymes that break glycosidic bonds through a hydrolytic mechanism, are presently classified into 123-amino acid sequence based families (2). ␣-Mannosidases known to process N-glycans are found in families 38, 47, 76, 92, and 99. Very recent studies have shown the bacterial family 38 ␣-mannosidase from Streptococcus pyogenes (SpyGH38) to be a specific exo-␣1,3-mannosidase that is tolerant of the ␣1,6-branches in N-glycans (3). Analysis of family 92 glycoside hydrolases from the human gut symbiont Bacteroides thetaiotaomicron revealed an expanded repertoire of ␣-mannosidases (4). These enzymes displayed activity primarily toward ␣1,2-and ␣1,3-mannosidic linkages with some having low ␣1,6-mannosidase activity. In addition to the established ability of S. pneumoniae to exo-hydrolytically process the distal arms of complex glycans, which comprise sialic acid, galactose, and N-acetylglucosamine, consideration of additional putative carbohydrate-active enzymes found in this organism suggests it can partly degrade the mannose component of N-glycans using enzymes similar to those found in S. pyogenes and B. thetaiotaomicron. Through these observations it has become clear that some bacteria, possibly including S. pneumoniae, have the capacity to process the mannose component of N-glycans. A noteworthy gap, however, in the known bacterial N-glycan de...

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“…This label facilitated sensitive detection by excitation with the stable argon-ion or solid state 488 nm lasers commonly used in commercial CE equipments, also providing good separation characteristics [7]. The detection limit of laser-induced fluorescence (LIF) of APTS labeled sugars in CE is in the low femtomolar range [8]. The most widely applied reducing agents for these reductive amination reactions are sodium cyanoborohydride and 2-picoline borane.…”

Section: Introductionmentioning

confidence: 99%

A novel carbohydrate labeling method utilizing transfer hydrogenation-mediated reductive amination

Kovács

Papp

Horváth

et al. 2017

Journal of Pharmaceutical and Biomedical Analysis

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One of the most frequently used high-resolution glycan analysis methods in the biopharmaceutical and biomedical fields is capillary electrophoresis with laser-induced fluorescence (CE-LIF) detection. Glycans are usually labeled by reductive amination with a charged fluorophore containing a primary amine, which reacts with the aldehyde group at the reducing end of the glycan structures. In this reaction, first a Schiff base is formed that is reduced to form a stable conjugate by a hydrogenation reagent, such as sodium cyanoborohydride. In large scale biopharmaceutical applications, such as clone selection for glycoprotein therapeutics, hundreds of reactions are accomplished simultaneously, so the HCN generated in the process poses a safety concern. To alleviate this issue, here we propose catalytic hydrogen transfer from formic acid catalyzed by water-soluble iridium(III)-and ruthenium(II)-phosphine complexes as a novel alternative to hydrogenation. The easily synthesized water-soluble iridium(III) and the ruthenium(II) hydrido complexes showed high catalytic activity in carbohydrate labeling. This procedure is environmentally friendly and reduces the health risks for the industry. Using carbohydrate standards, oligosaccharides released from glycoproteins with highly sialylated (fetuin), high mannose (ribonuclease B) and mixed sialo and neutral (human plasma) N-glycans, we demonstrated similar labeling efficiencies for iridium(III) dihydride to that of the conventionally used sodium cyanoborohydride based reaction. The derivatization reaction time was less than 20 min with no bias towards the above mentioned specific glycan structures.

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Acid‐catalyzed reductive amination of aldoses with 8‐aminopyrene‐1,3,6‐trisulfonate

Cited by 57 publications

References 18 publications

Evaporative fluorophore labeling of carbohydrates via reductive amination

Evaporative fluorophore labeling of carbohydrates via reductive amination

Analysis of a New Family of Widely Distributed Metal-independent α-Mannosidases Provides Unique Insight into the Processing of N-Linked Glycans

A novel carbohydrate labeling method utilizing transfer hydrogenation-mediated reductive amination

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