Time-resolved fluorometric analysis of carbohydrates labeled with amino-aromatic compounds by reductive amination

Nakajima, Kazuki; Oda, Yasuo; Kinoshita, Mitsuhiro; Masuko, Takashi; Kakehi, Kazuaki

doi:10.1039/b202950b

Cited by 9 publications

(4 citation statements)

References 19 publications

(23 reference statements)

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“…Also, affinity CE (ACE) has been successfully applied to study the binding of ligands for receptors and some examples of the application of the technique to the screening of chemical libraries are reported [27][28][29][30]. The choice of the methodology has to take into account the characteristics of the receptor and of the ligand, the type of information desired, as well as the throughput needed.…”

Section: Introductionmentioning

confidence: 99%

Search of ligands for the amyloidogenic protein β2-microglobulin by capillary electrophoresis and other techniques

et al. 2005

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Beta2-microglobulin (beta2-m) is a small amyloidogenic protein normally present on the surface of most nucleated cells and responsible for dialysis-related amyloidosis, which represents a severe complication of long-term hemodialysis. A therapeutic approach for this amyloidosis could be based on the stabilization of beta2-m through the binding to a small molecule, and consequent inhibition of protein misfolding and amyloid fibril formation. A few compounds have been described to weakly bind beta2-m, including the drug suramin. The lack of a binding site for nonpolypeptidic ligands on the beta2-m structure makes it difficult for both the identification of functional groups responsible for the binding and the search of hits to be optimized. The characterization of the binding properties of suramin for beta2-m by using three different techniques (surface plasmon resonance, affinity CE (ACE), ultrafiltration) is here described and the results obtained are compared. The common features of the chemical structures of the compounds known to bind the protein led us to select 200 sulfonated/suramin-like molecules from a wider chemical library on the basis of similarity rules, so as to possibly single out some interesting hits and to gain more information on the functional groups involved in the binding. The development of screening methods to test the compounds by using ultrafiltration and ACE is described.

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Section: Introductionmentioning

confidence: 99%

Search of ligands for the amyloidogenic protein β2-microglobulin by capillary electrophoresis and other techniques

et al. 2005

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“…Preparation of standard samples of highly purified free oligosaccharides is quite important, because these oligosaccharides are easily converted to various forms for the studies such as carbohydrate-protein interactions. − If pure oligosaccharides can be obtained from natural source, then the library of oligosaccharides is useful for construction of sugar array, which offers a possibility for high-throughput analysis of interactions between carbohydrates and proteins (i.e., ‘glycome') . On the basis of these considerations, many challenges of chemical, chemoenzymatic, and biological synthesis of oligosaccharide have been reported. − Although these strategies are useful for the synthesis of target carbohydrates in mg-quantities or more, they are problematic in the preparation of diverse structures of oligosaccharides because they require multistep reactions which cause much labor.…”

Section: Introductionmentioning

confidence: 99%

Rapid and Sensitive Screening of N-Glycans as 9-Fluorenylmethyl Derivatives by High-Performance Liquid Chromatography: A Method Which Can Recover Free Oligosaccharides after Analysis

et al. 2005

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There are a large number of labeling methods for asparagine-type oligosaccharides with fluorogenic and chromophoric reagents. We have to choose the most appropriate labeling method based on the purposes such as mass spectrometry, high-performance liquid chromatography and capillary electrophoresis. Asparagine-type glycans are released from core proteins as N-glycosylamine at the initial step of the releasing reaction when glycoamidase F is employed as the enzyme. The N-glycosylamine-type oligosaccharides thus released by the enzyme are subjected to hydrolysis or mutarotation to form free-form oligosaccharides. In the detailed studies on the enzyme reaction, we found a condition in which the released N-glycosylamine-type oligosaccharides were exclusively present at least during the course of enzyme reaction, and developed a method for in situ derivatization of the glycosylamine-type oligosaccharides with 9-fluorenylmethyl chloroformate (Fmoc-Cl). The Fmoc labeled sialo- and asialo- (or high-mannose and hybrid) oligosaccharides were successfully analyzed on an amine-bonded polymer column and amide-silica column, respectively. The present method showed approximately 5 times higher sensitivities than that using 2-aminobenzoic acid (2-AA). The separation profile was similar to that observed using 2-AA method as examined by the analyses of carbohydrate chains derived from several glycoproteins including complex-type, high-mannose type and hybrid type of N-linked oligosaccharides. The labeled oligosaccharides were stable at least for several months when stored at -20 degrees C. Furthermore, it should be emphasized that the Fmoc-derivatized oligosaccharides could be easily recovered as free reducing oligosaccharides simply by incubation with morpholine in dimethylformamide solution. We obtained a pure triantennary oligosaccharide with 3 sialic acid residues as a free reducing form from fetuin in good yield after isolation of the corresponding Fmoc oligosaccharide followed by removing reaction of the Fmoc group. The proposed method will be useful for preparation of free oligosaccharides as standard samples at pmol-nmol scale from commercially available glycoproteins.

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“…An additional function of these labeling reagents is to improve ionization efficiency and glycan stability in both matrix-assisted laser desorption ionization (MALDI) and electrospray ionization (ESI). Several other labeling reagents have also been reported for quantitative glycomics, including 2-aminoacridone (AMAC) (Charlwood, Birrell, Organ, & Camilleri, 1999), 4-aminobenzonitrile (ABN) (Schwaiger, Oefner, Huber, Grill, & Bonn, 1994), 3-aminobenzoic acid (3-AA) (Nakajima, Oda, Kinoshita, Masuko, & Kakehi, 2002) and 3-aminobenzamide (3-AB) (Kakehi, Funakubo, Suzuki, Oda, & Kitada, 1999). …”

Section: 2 Sample Handling In Glycomicsmentioning

confidence: 99%

Quantitative Glycomics

Veillon

Zhou

Mechref

2017

Methods in Enzymology

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Glycosylation is one of the most common and essential protein modifications. Glycans conjugated to biomolecules modulate the function of such molecules through both direct recognition of glycan structures and indirect mechanisms that involve the control of protein turnover rates, stability, and conformation. The biological attributes of glycans in numerous biological processes and implications in a number of diseases highlight the necessity for comprehensive characterization of protein glycosylation. This chapter review cutting-edge-methods and tools developed to facilitated quantitative glycomics. The chapter highlight the different methods employed for the release and purification of glycans from biological samples. The most effective labeling methods developed for sensitive quantitative glycomics are also described and discussed. The chromatographic approaches that have been used effectively in glycomics are also highlighted.

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Time-resolved fluorometric analysis of carbohydrates labeled with amino-aromatic compounds by reductive amination

Cited by 9 publications

References 19 publications

Search of ligands for the amyloidogenic protein β2-microglobulin by capillary electrophoresis and other techniques

Search of ligands for the amyloidogenic protein β2-microglobulin by capillary electrophoresis and other techniques

Rapid and Sensitive Screening of N-Glycans as 9-Fluorenylmethyl Derivatives by High-Performance Liquid Chromatography: A Method Which Can Recover Free Oligosaccharides after Analysis

Quantitative Glycomics

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