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Site-specific analysis of advanced glycation end products in plasma proteins of type 2 diabetes mellitus patients

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Abstract

Advanced glycation end products (AGEs) are posttranslational modifications formed non-enzymatically from the reaction of carbohydrates and their degradation products with proteins. Accumulation of AGEs is associated with the progression of severe diabetic complications, for example, and elevated tissue levels of AGEs might even predict these pathologies. As AGE formation is often site-specific, mapping of these modification sites may reveal more sensitive and specific markers than the global tissue level. Here, 42 AGE modifications were identified in a bottom-up proteomic approach by tandem mass spectrometry, which corresponded to 36 sites in 22 high to medium abundant proteins in individual plasma samples obtained from type 2 diabetes mellitus (T2DM) patients with long disease duration (>10 years). Major modifications were glarg (11 modification sites) and carboxymethylation (5) of arginine and formylation (8), acetylation (7), and carboxymethylation (7) of lysine residues. Relative quantification of these sites in plasma samples obtained from normoglycemic individuals (n = 47) and patients with T2DM being newly diagnosed (n = 47) or of medium (2–5 years, n = 20) and long disease duration (>10 years, n = 20) did not reveal any significant differences.

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Acknowledgments

The authors thank Sandro Spiller and Dr. Ravi Chand Bollineni for assisting parallel sample processing. Funding from the Deutsche Forschungsgemeinschaft (DFG, grant number HO-2222/7-1 to RH) and a PhD scholarship from the Ernst Schering Foundation to UG are gratefully acknowledged.

Author contributions

Uta Greifenhagen conducted the experiments, data analysis, and interpretation and wrote the manuscript together with Ralf Hoffmann. Andrej Frolov contributed to sample preparation and the experimental design. Matthias Blüher designed the cohorts and provided the plasma samples. Ralf Hoffmann initiated the project, contributed to data interpretation, and wrote the manuscript together with Uta Greifenhagen. All the authors reviewed and agreed upon the final version of this manuscript.

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Author notes

  1. Andrej Frolov

    Present address: Department of Bioorganic Chemistry, Leibniz Institute of Plant Biochemistry, Weinberg 3, 06120, Halle, Germany

Authors and Affiliations

  1. Institute of Bioanalytical Chemistry, Faculty of Chemistry and Mineralogy, Universität Leipzig, Deutscher Platz 5, 04103, Leipzig, Germany

    Uta Greifenhagen, Andrej Frolov & Ralf Hoffmann

  2. Center for Biotechnology and Biomedicine (BBZ), Universität Leipzig, Deutscher Platz 5, 04103, Leipzig, Germany

    Uta Greifenhagen & Ralf Hoffmann

  3. Endocrinology, Department of Medicine, Universität Leipzig, Liebigstraße 20, 04103, Leipzig, Germany

    Matthias Blüher

Authors
  1. Uta Greifenhagen
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Corresponding author

Correspondence to Ralf Hoffmann.

Ethics declarations

The study was approved by the Ethics Committee of Universität Leipzig (approval no: 159-12-21052012) and performed in accordance with the Declaration of Helsinki. All the subjects gave written informed consent before taking part in this study.

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The authors declare that they have no conflict of interest.

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Greifenhagen, U., Frolov, A., Blüher, M. et al. Site-specific analysis of advanced glycation end products in plasma proteins of type 2 diabetes mellitus patients. Anal Bioanal Chem 408, 5557–5566 (2016). https://doi.org/10.1007/s00216-016-9651-4

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  • DOI: https://doi.org/10.1007/s00216-016-9651-4

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