Glycation and oxidation of histones H2B and H1: in vitro study and characterization by mass spectrometry
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Among the post-translational modifications, oxidation and glycation are of special interest, especially in diseases such as diabetes, and in aging. The synergistic interaction between glycation and oxidation, also known as “glycoxidation” is highly relevant due to its involvement in the production of deleterious changes at the molecular level. Non-enzymatic damage to nuclear proteins has potentially severe consequences for the maintenance of genomic integrity . In this report, we study glycated histones and its in vitro oxidation. Data concerning the modifications that occurred in the histones were obtained by analysis of enzymatic digests (Glu-C and Arg-C) of unmodified and glycated histones, obtained before and after oxidation. Analysis was then performed using a MALDI-MS/MS-based approach combined with nano liquid chromatography. This approach allowed us to identify histone H2B and H1 specific-sites of oxidation and to distinguish the most affected residues for each histone. The results showed the occurrence of a cumulative effect of oxidative damage in the glycated histones when subjected to in vitro oxidation, suggesting that structural changes caused by glycation induces histones to a pro-oxidant state. Comparing the data of oxidized glycated histones with data from unmodified oxidized histones, using the same model of oxidation, the results clearly show that these oxidative modifications occur earlier and more extensively in glycated histones. Furthermore, the results pointed to an increased oxidative damage in the vicinity of the glycated residues.
KeywordsHistones Glycation Protein oxidation Mass spectrometry
The authors express their appreciation for the financial support provided by the “Fundação para a Ciência e Tecnologia” (Grant no SFRH/BD/14963/2004 and PTDC/QUI/68408/2006).
- 15.Amadori M (1925) Atii real accad naz Lincei 2:337–345Google Scholar
- 17.Wolff SP, Dean RT (1987) Glucose autoxidation and protein modification. The potential role of ‘autoxidative glycosylation’ in diabetes. Biochem J 245:243–250Google Scholar
- 31.Bonenfant D, Coulot M, Towbin H, Schindler P, van Oostrum J (2006) Characterization of histone H2A and H2B variants and their post-translational modifications by mass spectrometry. Mol Cell Proteomics 5:541–552Google Scholar
- 37.Zhang K, Tang H, Huang L, Blankenship JW, Jones PR, Xiang F, Yau PM, Burlingame AL (2002) Identification of acetylation and methylation sites of histone H3 from chicken erythrocytes by high-accuracy matrix-assisted laser desorption ionization-time-of-flight, matrix-assisted laser desorption ionization-postsource decay, and nanoelectrospray ionization tandem mass spectrometry. Anal Biochem 306:259–269CrossRefGoogle Scholar
- 50.Stadtman ER, Berlett BS (1991) Fenton chemistry. Amino acid oxidation. J Biol Chem 266:17201–17211Google Scholar
- 54.Wondrak GT, Cervantes-Laueant, Jacobson EL, Jacobson MK (2000) Histone carbonylation in vivo and in vitro. Biochem. J. 351:769–777Google Scholar