“Zoom-In”—A Targeted Database Search for Identification of Glycation Modifications Analyzed by Untargeted Tandem Mass Spectrometry

Bhonsle, Hemangi S.; Korwar, Arvind M.; Kesavan, Suresh K.; Bhosale, Santosh D.; Bansode, Sneha; Kulkarni, Mahesh J.

doi:10.1255/ejms.1203

Cited by 13 publications

(11 citation statements)

References 30 publications

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“…The search-algorithm-identified glycated peptides were manually validated by inspecting each MS/MS spectrum as described earlier by (27). Briefly, at MS, each AGE-modified peptide precursor was reviewed for presence of unmodified peptide precursor.…”

Section: Resultsmentioning

confidence: 99%

“…A capillary voltage of 3 kV, source temperature of 80°C, and cone voltage of 32 V were maintained during the analyses. LC-MSE data were processed using Protein Lynx Global Server (PLGS, Version 2.5.1) for identification and quantification of glycated peptides of albumin (27,35).…”

Section: Targeted Quantification Of Glycated Peptides Of Hsa In Diabementioning

confidence: 99%

“…Further, by using a combination of immunodepletion, enrichment and fractionation strategies, a total of 7,749 unique glycated peptides corresponding to 1,095 native human plasma proteins, 1,592 in vitro glycated human plasma proteins, and 1,664 erythrocyte proteins were identified (26). In these lines, we have previously reported a database search approach for the identification of glycated peptide in a crude or nonenriched sample by untargeted MS/MS or data-independent workflow (27). Glycation is chronic process; a given protein can undergo dynamic heterogeneous transformations as these proteins have varying biological lifespans, influencing the function of a protein.…”

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confidence: 99%

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Development of Diagnostic Fragment Ion Library for Glycated Peptides of Human Serum Albumin: Targeted Quantification in Prediabetic, Diabetic, and Microalbuminuria Plasma by Parallel Reaction Monitoring, SWATH, and MSE

Korwar

Vannuruswamy

Jagadeeshaprasad

et al. 2015

Molecular & Cellular Proteomics

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Human serum albumin is one of the most abundant plasma proteins that readily undergoes glycation, thus glycated albumin has been suggested as an additional marker for monitoring glycemic status. Hitherto, only Amadori-modified peptides of albumin were quantified. In this study, we report the construction of fragment ion library for Amadori-modified lysine (AML), N()-(carboxymethyl)lysine (CML)-, and N()-(carboxyethyl)lysine (CEL)-modified peptides of the corresponding synthetically modified albumin using high resolution accurate mass spectrometry (HR/AM). The glycated peptides were manually inspected and validated for their modification. Further, the fragment ion library was used for quantification of glycated peptides of albumin in the context of diabetes. Targeted Sequential Window Acquisition of all THeoretical Mass Spectra (SWATH) analysis in pooled plasma samples of control, prediabetes, diabetes, and microalbuminuria, has led to identification and quantification of 13 glycated peptides comprised of four AML, seven CML, and two CEL modifications, representing nine lysine sites of albumin. Five lysine sites namely K549, K438, K490, K88, and K375, were observed to be highly sensitive for glycation modification as their respective m/z showed maximum fold change and had both AML and CML modifications. Thus, peptides involving these lysine sites could be potential novel markers to assess the degree of glycation in diabetes. Molecular & Cellular Proteomics

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Section: Resultsmentioning

confidence: 99%

Section: Targeted Quantification Of Glycated Peptides Of Hsa In Diabementioning

confidence: 99%

mentioning

confidence: 99%

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Development of Diagnostic Fragment Ion Library for Glycated Peptides of Human Serum Albumin: Targeted Quantification in Prediabetic, Diabetic, and Microalbuminuria Plasma by Parallel Reaction Monitoring, SWATH, and MSE

Korwar

Vannuruswamy

Jagadeeshaprasad

et al. 2015

Molecular & Cellular Proteomics

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“…E -AGE modification analysis for human serum albumin (HSA) from clinical CICs was performed as described earlier (18). Briefly, a targeted search was performed involving variable glycation modifications, namely amadori (ϩ162.0528) at lysine or arginine, carboxymethyl lysine (ϩ58.0055) and carboxyethyl lysine (ϩ72.0211) at lysine, methyl-glyoxal-induced hydroimidazolone-1 (MGH1) (ϩ 54.0106) at arginine, and oxidation at Met and fixed carbamidomethylation of Cys residues.…”

Section: Age Modification Analysis By Lc-msmentioning

confidence: 99%

Proteomic Insight Reveals Elevated Levels of Albumin in Circulating Immune Complexes in Diabetic Plasma

Bhat

Jagadeeshaprasad

Patil

et al. 2016

Molecular & Cellular Proteomics

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A Hyperglycemic condition in diabetes promotes formation of advanced glycation end products, which are known to elicit immune response and form complexes with immunoglobulins called circulating immune complexes. To investigate the involvement of advanced glycation end product (AGE)-modified proteins in the elicitation of an immune response, circulating immune complexes were isolated and proteins associated were identified and characterized. Label-free-based mass spectrometric analysis of circulating immune complexes in clinical plasma of prediabetic, newly diagnosed diabetes, and diabetic microalbuminurea revealed elevated levels of serum albumin in the circulating immune complexes, which were also observed to be AGE modified. Further, to examine the role of glycation, circulating immune complexes were analyzed in the streptozotocin-induced diabetic mice treated with or without aminoguanidine, a prototype glycation inhibitor. Mass spectrometric analysis of circulating immune complexes showed elevated levels of serum albumin in plasma from diabetic mice over that of control animals. Aminoguanidine-treated diabetic mice displayed decreased AGE modification of plasma albumin, accompanied by a reduced level of albumin in the circulating immune complexes. In addition, elevated levels of proinflammatory cytokines such as IL-1b, IL-2, and TNF-alpha were observed in diabetes, which were reduced with aminoguanidine treatment, suggesting the involvement of glycation in the immune response.

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“…Recent advances in mass spectrometry have led to more definitive identifications of AGEs associated with proteins [19; 20; 21; 22; 23; 24]. However, limited application of mass spectrometry to localize the precursors to AGEs, the Amadori products, have been reported [22; 25].…”

Section: Introductionmentioning

confidence: 99%

Identification and characterization of glycation adducts on osteocalcin

Thomas

Cleland

Zhang

et al. 2017

Analytical Biochemistry

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Osteocalcin is an important extracellular matrix bone protein that contributes to the structural properties of bone through its interactions with hydroxyapatite mineral and with collagen I. This role may be affected by glycation, a labile modification the levels of which has been shown to correlate with bone fragility. Glycation starts with the spontaneous addition of a sugar onto a free amine group on a protein, forming an Amadori product, and then proceeds through several environment-dependent stages resulting in the formation of an advanced glycation end product. Here, we induce the first step of this modification on synthetic osteocalcin, and then use multiple mass spectrometry fragmentation techniques to determine the location of this modification. Collision-induced dissociation resulted in spectra dominated by neutral loss, and was unable to identify Amadori products. Electron-transfer dissociation showed that the Amadori product formed solely on osteocalcin’s N-terminus. This suggests that the glycation of osteocalcin is unlikely to interfere with osteocalcin’s interaction with hydroxyapatite. Instead, glycation may interfere with its interaction with collagen I or another bone protein, osteopontin. Potentially, the levels of glycated osteocalcin fragments released from bone during bone resorption could be used to assess bone quality, should the N-terminal fragments be targeted.

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“Zoom-In”—A Targeted Database Search for Identification of Glycation Modifications Analyzed by Untargeted Tandem Mass Spectrometry

Cited by 13 publications

References 30 publications

Development of Diagnostic Fragment Ion Library for Glycated Peptides of Human Serum Albumin: Targeted Quantification in Prediabetic, Diabetic, and Microalbuminuria Plasma by Parallel Reaction Monitoring, SWATH, and MSE

Development of Diagnostic Fragment Ion Library for Glycated Peptides of Human Serum Albumin: Targeted Quantification in Prediabetic, Diabetic, and Microalbuminuria Plasma by Parallel Reaction Monitoring, SWATH, and MSE

Proteomic Insight Reveals Elevated Levels of Albumin in Circulating Immune Complexes in Diabetic Plasma

Identification and characterization of glycation adducts on osteocalcin

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