The efficient synthesis of isotopically labeled peptide-derived Amadori products and their characterization

Kapczyńska, Katarzyna; Stefanowicz, Piotr; Jaremko, Łukasz; Jaremko, Mariusz; Kluczyk, Alicja; Szewczuk, Zbigniew

doi:10.1007/s00726-010-0714-0

Cited by 9 publications

(5 citation statements)

References 28 publications

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“…This isotopic exchange was previously detected by Heyns and co‐workers14 who noted that signals of protons at the position C‐1 in Amadori rearrangement compounds decreased on prolonged storage of solutions in D 2 O because of H/D exchange, which significantly accelerated with increasing pH values15 (see the Supporting Information).…”

Section: Resultssupporting

confidence: 76%

The Amadori Rearrangement for Carbohydrate Conjugation: Scope and Limitations

et al. 2016

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The Amadori rearrangement was investigated for the synthesis of C‐glycosyl‐type neoglycoconjugates. Various amines including diamines, amino‐functionalized glycosides, lysine derivatives, and peptides were conjugated with two different heptoses to generate non‐natural C‐glycosyl‐type glycoconjugates of the d‐gluco and d‐manno series. With these studies, the scope and limitations of the Amadori rearrangement as a conjugation method have been exemplified with respect to the carbohydrate substrate, as well as the amino components.

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Section: Resultssupporting

confidence: 76%

The Amadori Rearrangement for Carbohydrate Conjugation: Scope and Limitations

et al. 2016

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“…The signals of the α-and β-furanose forms could be clearly assigned too, but their intensity is a factor 3-6 less intense in agreement to the previously reported abundances [26,44]. Further comparison with assignments of Nα-(1-deoxy-d-fructos-1-yl)-l-alanine [27] and a peptide in which an N-terminal Lys was glycated at its side chain, [41] confirm these assignments to the different forms. There are small differences in the previously reported values, but this is likely due to the slightly different structures of the glycated moieties.…”

Section: Analysis Of Artificially Glycated Model Proteins Under Denat...supporting

confidence: 88%

“…Although Amadori products were analyzed before using model compounds [ 26 , 41 ], glycation of proteins has not been investigated by NMR spectroscopy so far. To obtain a sample with glycated lysine(s), we incubated protein and peptide solutions in the presence of phosphate with high concentrations of glucose at 40°C instead of using solid phase peptide synthesis using glycated precursors [ 41 , 42 ]. Initial attempts with short peptides containing lysine and arginine gave only poor yields.…”

Section: Resultsmentioning

confidence: 99%

“…S1C, D). As the first set of chemical shifts was most similar to the ones observed in the other glycated proteins, we (26,27,41,45) a The two columns represent the two detected spin systems b Values of compound 5: Nα-(1-deoxy-d-fructos-1-yl)-l-alanine. For comparison with our data referenced to DSS, we added + 2.5 ppm to the values of Kaufmann referenced to TMS c Values of the peptide H-Lys([ 13 C 6 ]Fru)-Ala-Ala-Phe-OH d Values of compound 6 Nϵ-(1-deoxy-d-fructos-1-yl)-Nα-formyl-l-lysine.…”

Section: Analysis Of Artificially Glycated Model Proteins Under Denat...mentioning

confidence: 85%

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Unambiguous Identification of Glucose-Induced Glycation in mAbs and other Proteins by NMR Spectroscopy

et al. 2022

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Objective Glycation is a non-enzymatic and spontaneous post-translational modification (PTM) generated by the reaction between reducing sugars and primary amine groups within proteins. Because glycation can alter the properties of proteins, it is a critical quality attribute of therapeutic monoclonal antibodies (mAbs) and should therefore be carefully monitored. The most abundant product of glycation is formed by glucose and lysine side chains resulting in fructoselysine after Amadori rearrangement. In proteomics, which routinely uses a combination of chromatography and mass spectrometry to analyze PTMs, there is no straight-forward way to distinguish between glycation products of a reducing monosaccharide and an additional hexose within a glycan, since both lead to a mass difference of 162 Da. Methods To verify that the observed mass change is indeed a glycation product, we developed an approach based on 2D NMR spectroscopy spectroscopy and full-length protein samples denatured using high concentrations of deuterated urea. Results The dominating β-pyranose form of the Amadori product shows a characteristic chemical shift correlation pattern in 1H-13C HSQC spectra suited to identify glucose-induced glycation. The same pattern was observed in spectra of a variety of artificially glycated proteins, including two mAbs, as well as natural proteins. Conclusion Based on this unique correlation pattern, 2D NMR spectroscopy can be used to unambiguously identify glucose-induced glycation in any protein of interest. We provide a robust method that is orthogonal to MS-based methods and can also be used for cross-validation.

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“…13%; and α-furanose: ca. 12%) [38,39]. Boronic acid is known to bind glucose preferentially in the α-furanose form and not in the more abundant α-pyranose form.…”

Section: Discussionmentioning

confidence: 99%

An Optimised Di-Boronate-ChemMatrix Affinity Chromatography to Trap Deoxyfructosylated Peptides as Biomarkers of Glycation

et al. 2020

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We report herein a novel ChemMatrix® Rink resin functionalised with two phenylboronate (PhB) moieties linked on the N-α and N-ε amino functions of a lysine residue to specifically capture deoxyfructosylated peptides, compared to differently glycosylated peptides in complex mixtures. The new PhB-Lys(PhB)-ChemMatrix® Rink resin allows for exploitation of the previously demonstrated ability of cis diols to form phenylboronic esters. The optimised capturing and cleavage procedure from the novel functionalised resin showed that only the peptides containing deoxyfructosyl-lysine moieties can be efficiently and specifically detected by HR-MS and MS/MS experiments. We also investigated the high-selective affinity to deoxyfructosylated peptides in an ad hoc mixture containing unique synthetic non-modified peptides and in the hydrolysates of human and bovine serum albumin as complex peptide mixtures. We demonstrated that the deoxyfructopyranosyl moiety on lysine residues is crucial in the capturing reaction. Therefore, the novel specifically-designed PhB-Lys(PhB)-ChemMatrix® Rink resin, which has the highest affinity to deoxyfructosylated peptides, is a candidate to quantitatively separate early glycation peptides from complex mixtures to investigate their role in diabetes complications in the clinics.

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The efficient synthesis of isotopically labeled peptide-derived Amadori products and their characterization

Cited by 9 publications

References 28 publications

The Amadori Rearrangement for Carbohydrate Conjugation: Scope and Limitations

The Amadori Rearrangement for Carbohydrate Conjugation: Scope and Limitations

Unambiguous Identification of Glucose-Induced Glycation in mAbs and other Proteins by NMR Spectroscopy

An Optimised Di-Boronate-ChemMatrix Affinity Chromatography to Trap Deoxyfructosylated Peptides as Biomarkers of Glycation

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