A study on in vitro glycation processes by matrix-assisted laser desorption ionization mass spectrometry

Lapolla, Annunziata; Gerhardinger, Chiara; Baldo, L.; Fedele, Domenico; Keane, Anita; Seraglia, Roberta; Catinella, Silvia; Traldi, Pietro

doi:10.1016/0925-4439(93)90118-k

Cited by 49 publications

(26 citation statements)

References 25 publications

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“…In order to verify the MALDI effectiveness in the ®eld of interest, some preliminary studies were carried out on in vitro glycated proteins. The ®rst experiments involved the incubation of BSA in pseudophysiological conditions with glucose at various concentrations (Lapolla et al, 1993). Samples were drawn at differing incubation times and, after extensive dialysis to eliminate free glucose and further lyophilization, samples were analyzed by MALDI.…”

Section: Evaluation Of Glycation Of Native Proteinsmentioning

confidence: 99%

The role of mass spectrometry in the study of non-enzymatic protein glycation in diabetes

Lapolla¹,

Fedele²,

Traldi³

2000

Mass Spectrom. Rev.

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Section: Evaluation Of Glycation Of Native Proteinsmentioning

confidence: 99%

The role of mass spectrometry in the study of non-enzymatic protein glycation in diabetes

Lapolla¹,

Fedele²,

Traldi³

2000

Mass Spectrom. Rev.

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“…For all of these experiments, the instrument was set at linear mode to increase the mass range and sensitivity. To ensure mass accuracy, an external calibration was performed prior to each sample spot, and a calibration check was performed after each sample to ensure that the calibration had not shifted (see supplemental material) (43,44).…”

Section: Methodsmentioning

confidence: 99%

Paradoxical Impact of Antioxidants on Post-Amadori Glycoxidation

Culbertson

Vassilenko

Morrison

et al. 2003

Journal of Biological Chemistry

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The inhibition of post-Amadori advanced glycation end product (AGE) formation by three different classes of AGE inhibitors, carbonyl group traps, chelators, and radical-trapping antioxidants, challenge the current paradigms that: 1) AGE inhibitors will not increase the formation of any AGE product, 2) transition metal ions are required for oxidative formation of AGE, and 3) screening AGE inhibitors only in systems containing transition metal ions represents a valid estimate of potential in vivo mechanisms. This work also introduces a novel multifunctional AGE inhibitor, 6-dimethylaminopyridoxamine (dmaPM), designed to function as a combined carbonyl trap, metal ion chelator, and radicaltrapping antioxidant. Other AGE inhibitors including pyridoxamine, aminoguanidine, o-phenylenediamine, dipyridoxylamine, and diethylenetriaminepentaacetic acid were also examined. The results during uninterrupted and interrupted ribose glycations show: 1) an unexpected increase in the yield of pentosidine in the presence of radical-trapping phenolic antioxidants such as Trolox and dmaPM, 2) significant formation of N ⑀ -carboxymethyllysine (CML) in the presence of strong chelators and phenolic antioxidants, which implies that there must be nonradical routes to CML, 3) prevention of intermolecular cross-links with radical-trapping inhibitors, and 4) that dmaPM shows excellent inhibition of AGE. Glucose glycations reveal the expected inhibition of pentosidine and CML with all compounds tested, but in a buffer free of trace metal ions the yield of CML in the presence of radical-trapping antioxidants was between the metal ion-free and metal ion-containing controls. Protein molecular weight analyses support the conclusion that Amadori decomposition pathways are constrained in the presence of metal ion chelators and radical traps.Nonenzymatic protein glycation by reducing sugars, such as glucose or ribose, is a complicated cascade of condensations, rearrangements, fragmentations, and oxidative modifications that lead to a plethora of compounds collectively called advanced glycation end products (AGEs) 1 (1). AGE products slowly build up and contribute to the age-dependent chemical modification of long-lived tissue proteins. Once formed, AGE products may prevent normal protein function and recognition or stimulate potentially detrimental interactions in signaling pathways (2, 3). Glycation reactions are believed to have a significant role in the progression of diabetic complications largely because of elevated levels of glucose. Indeed, the formation of AGE has been increasingly implicated in pathogenesis of diabetic complications, atherosclerosis, and Alzheimer's disease (4 -6). Inhibitors that act to reduce AGE formation may prove useful for limiting nonenzymatic protein modification associated in the pathology of these and other chronic agerelated diseases. The reaction of amino compounds, such as lysine, with reducing sugars is known as the Maillard reaction (7, 8). The "classical" or Hodge pathway begins with reversible formation of...

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“…In previous investigations on protein glycation, under both in vitro 11,12 and in vivo [13][14][15] conditions, the MALDI 16 technique has revealed itself to be a valuable tool. Its sensitivity, specificity and mass accuracy not only yield the number of glucose molecules condensing on a specific protein, but also supply information on the dehydration/ oxidation levels of the glycated products.…”

mentioning

confidence: 98%

Direct evaluation of glycated and glyco-oxidized globins by matrix-assisted laser desorption/ionization mass spectrometry

Lapolla¹,

Fedele²,

Plebani³

et al. 1999

Rapid Commun. Mass Spectrom.

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Matrix-assisted laser desorption ionization (MALDI) mass spectrometry (MS) has been applied to achieve a qualitative and quantitative evaluation of glycated globins on a wide number of healthy and diabetic subjects. The method allowed us to establish that both alpha- and beta-globins are glycated and that, in addition to simply glycated products, other species are detected. Investigations by different sample treatments and by analysis of the glycated beta-globin fraction obtained by preparative chromatography indicated that these species correspond to glyco-oxidized globins. Consequently MALDI-MS can be validly employed to evaluate not only the glycation level, but also the degree of oxidative stress. The percentages of glycated and glycooxidized species were calculated from the related MALDI spectra by the measurement of the related peak areas, without any other treatment of data. A linear relationship between HbA1c values and the total percentage of glycated and glyco-oxidized globins has been found, and its slope (< 1) has been rationalized by the uncorrected evaluation of glycated globins content in the standard samples employed for HbA1c measurements.

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A study on in vitro glycation processes by matrix-assisted laser desorption ionization mass spectrometry

Cited by 49 publications

References 25 publications

The role of mass spectrometry in the study of non-enzymatic protein glycation in diabetes

The role of mass spectrometry in the study of non-enzymatic protein glycation in diabetes

Paradoxical Impact of Antioxidants on Post-Amadori Glycoxidation

Direct evaluation of glycated and glyco-oxidized globins by matrix-assisted laser desorption/ionization mass spectrometry

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