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Poster De Conférence Année : 2022

Chiral Amino Acid and Peptide Separations for Proteomics Applied to Aging Collagens

Résumé

Recent advances in LC-MS separation science have made it possible to separate chiral compounds, i.e. amino acids [1]. In nature, amino acids in proteins are found in their L-form. However, during the animal aging process, including in humans, a natural amino acid racemization can take place. Indeed, the number of D-amino acids in aging proteins has been shown to correlate with age [2]. D-amino acids in aging proteins, like D-Asp in elastin, β-amyloid, and α-crystallin, are linked to aging diseases i.e., arteriosclerosis [2], Alzheimer’s disease [3] and cataract [4]. For this work, the D-amino acids rates in recent and aging collagens at different ages were studied as well as the aging degradation of collagen sequences using peptidomics. Successive enzymatic digestions of collagens and arising peptide separations allow the collecting of these peptides into different fractions by HPLC. Peptides in each fraction were separated and identified by nanoLC-MS. After hydrolysis and derivatization, individual fractions were analyzed for proteinogenic D- and L-amino acid rates determination by a chiral LC-MS method. The % D-amino acid was progressive following age. Peptide mapping was used to determine the exact position of D-amino acids totally racemized. The comparison of peptide mapping between recent and aging collagen samples also made it possible to determine peptides resulting from enzymatic digestions and those resulting from aging-related degradations. The number of peptides that came from aging degradation increases according to the age of the samples. A novel chiral separation method by LC-MS was used for the enantiomeric determination of amino acids in collagen samples at different ages. A correlation between % D-amino acids and age was established. The exact positions of D-amino acids totally racemized were identified on the collagen sequences thanks to the peptide mapping. This peptide mapping was also used to determine peptides from enzymatic digestions and aging degradations. This analysis strategy can also be applied to collagen samples implicated in diseases such as osteogenesis imperfecta or several dysplasias, and all proteins related to aging and/or diseases naturally. [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-04319111 , version 1 (02-12-2023)

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  • HAL Id : hal-04319111 , version 1

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Marine Morvan, Ivan Mikšík. Chiral Amino Acid and Peptide Separations for Proteomics Applied to Aging Collagens. HPLC 2022, Jun 2022, San Diego, United States. ⟨hal-04319111⟩
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