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Study of Lysozyme Glycation Reaction by Mass Spectrometry and NMR Spectroscopy
Author(s) -
Maekawa Yukiko,
Sugiura Makiko,
Takeuchi Atsuko,
Tomoo Koji,
Ishida Toshimasa,
Kamigauchi Miyoko
Publication year - 2010
Publication title -
helvetica chimica acta
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.74
H-Index - 82
eISSN - 1522-2675
pISSN - 0018-019X
DOI - 10.1002/hlca.200900330
Subject(s) - lysozyme , glycation , chemistry , trimer , dimer , mass spectrometry , nuclear magnetic resonance spectroscopy , maillard reaction , glycosylation , proton nmr , chromatography , stereochemistry , biochemistry , organic chemistry , receptor
The Advanced Glycation End Products (AGEs) are the causative substances of lifestyle‐habit illness. To elucidate the glycation mechanism of the protein, the reaction of lysozyme with D ‐glucose was analyzed by the fluorescence, TOF‐MS, and 13 C‐NMR spectroscopy under the physiological condition. The fluorescence intensity of lysozyme in the glycation solution increased proportionally with a reaction time of ten weeks. The MALDI‐TOF‐MS spectra of the reaction solution after two weeks showed a peak at m / z 15066, which indicated the presence of a larger molecule than the native lysozyme ( m / z 14331), and new peaks at m / z 30105 (dimer) and 45000 (trimer) were also observed. The spectral analysis supported the assumption of a continuous glycation reaction of D ‐glucose with lysozyme and a 30% transformation of lysozyme to the dimeric form during ten weeks. The 13 C‐NMR spectra of lysozyme showed six [ 13 C]‐labeled signals by the glycation reaction with [ 13 C]‐glucose after two weeks of reaction. The combined analysis of TOF‐MS and 13 C‐NMR spectra uncovered that first products of the glycation reaction of lysozyme with D ‐glucose can be observed already three hours after starting the reaction and that nine D ‐glucose units are attached during ten weeks at 37°.