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Chiral amino acids analysis and aging proteomics applied to collagens

Résumé

Recent advances in LC-MS and CE-MS have allowed separating chiral compounds, i.e. amino acids [1]. In nature, proteinogenic amino acids are found in their L-form. However, a natural amino acid racemization can take place during the aging process. Indeed, the detection of D-amino acids in aging proteins has been shown to correlate with age [2] and linked to aging diseases. It is the case of D-Aps in elastin, β-amyloid and α-crystallin linked to arteriosclerosis [2], Alzheimer’s disease [3] and cataracts [4]. For these studies, peptides resulting from aging degradation of collagens at different ages were identified, and the D-amino acids rate and their position in aging collagen sequences were determined. After enzymatic digestion of collagen samples at different ages, the resulting peptides were separated and collected in several fractions. In each fraction, the peptides from aging collagens were compared to those of recent collagens. Peptides that are not common were identified as peptides resulting from age-related degradation. The number of these peptides increases with age. Then the peptide fractions were hydrolyzed and derivatized to obtain amino acid fractions. Finally, a chiral LC-MS separation method allows the determination of D-amino acid rates in fractions of aging collagen samples. Thanks to peptide mapping, the exact position of the fully racemized D-amino acids has been elucidated. This study made it possible to distinguish the peptides from enzymatic digestion and age-related degradations produce during the natural aging process. Then, a novel chiral separation method has allowed determining the % D-amino acid in collagen samples at different ages. A correlation between this % D-amino acid and age was confirmed. Finally, the peptide mapping has allowed identifying the exact position of D-amino acids totally racemized in the collagen sequence. This analysis strategy can also be applied to collagen samples implicated in diseases, and all proteins related to aging and/or diseases. [1] M. Morvan and I. Mikšík, Recent Advances in Chiral Analysis of Proteins and Peptides, Separations, 2021, 8, 112. [2] J. T. Powell, N. Vine, and M. Crossman, On the accumulation of d-aspartate in elastin and other proteins of the ageing aorta, Atherosclerosis, 1992, 97, 201–208. [3] J. M. Lee, L. Petrucelli, G. Fisher, S. Ramdath, J. Castillo, M. M. Di Fiore, A. D’Aniello, Evidence for D-Aspartyl-β-Amyloid Secretase Activity in Human Brain, J Neuropathol Exp Neurol, 2002, 61, 125–131. [4] M. Y. S. Hooi and R. J. W. Truscott, Racemisation and human cataract. d-Ser, d-Asp/Asn and d-Thr are higher in the lifelong proteins of cataract lenses than in age-matched normal lenses, Age, 2011, 33, 131–141.
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Dates et versions

hal-04319092 , version 1 (02-12-2023)

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  • HAL Id : hal-04319092 , version 1

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Marine Morvan, Ivan Mikšík. Chiral amino acids analysis and aging proteomics applied to collagens. Analytics 2022, Sep 2022, Nantes, France. ⟨hal-04319092⟩
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