Meta-proteomic analysis of two mammoth’s trunks by EVA technology and high-resolution mass spectrometry for an indirect picture of their habitat and the characterization of the collagen type I, alpha-1 and alpha-2 sequence

Abstract

The recent paleoproteomic studies, including paleo-metaproteomic analyses, improved our understanding of the dietary of ancient populations, the characterization of past human diseases, the reconstruction of the habitat of ancient species, but also provided new insights into the phylogenetic relationships between extant and extinct species. In this respect, the present work reports the results of the metaproteomic analysis performed on the middle part of a trunk, and on the portion of a trunk tip tissue of two different woolly mammoths some 30,000 years old. In particular, proteins were extracted by applying EVA (Ethylene–Vinyl Acetate studded with hydrophilic and hydrophobic resins) films to the surface of these tissues belonging to two Mammuthus primigenus specimens, discovered in two regions located in the Russian Far East, and then investigated via a shotgun MS-based approach. This approach allowed to obtain two interesting results: (i) an indirect description of the habitat of these two mammoths, and (ii) an improved characterization of the collagen type I, alpha-1 and alpha-2 chains (col1a1 and col1a2). Sequence characterization of the col1a1 and col1a2 highlighted some differences between M. primigenius and other Proboscidea together with the identification of three (two for col1a1, and one for col1a2) potentially diagnostic amino acidic mutations that could be used to reliably distinguish the Mammuthus primigenius with respect to the other two genera of elephantids (i.e., Elephas and Loxodonta), and the extinct American mastodon (i.e., Mammut americanum). The results were validated through the level of deamidation and other diagenetic chemical modifications of the sample peptides, which were used to discriminate the “original” endogenous peptides from contaminant ones. The data have been deposited to the ProteomeXchange with identifier < PXD029558 > .

A Peptidomics Method for Assessing Sex from Modern and Ancient Bovine Tooth Enamel.

“…Global deamidation and oxidation has been used as a tool to validate antiquity and study diagenesis in archaeological samples 32,34,35 . However, the average of the modifications across all sites in the protein/proteome reduces the sensitivity of the method.…”

Count Dracula Resurrected: Proteomic Analysis of Vlad III the Impaler’s Documents by EVA Technology and Mass Spectrometry

“…Synthesis of the EVA Films and Protein Harvesting. The synthesis of the EVA film was carried out under biologically and chemically clean conditions as previously reported by Cucina et al 20 In particular, a special plastic-like film based on EVA as a binder of ground AG 501 mix-bed cation/anion exchange, C8 and C18 resins (all from Bio-Rad) was prepared. The mixture was made comprising 70% 1−10 μm size ground beads and 30% EVA (the melting temperature was 75 °C).…”

Estrutura e composição química do esmalte dentário em vertebrados terrestres fósseis e recentes

“…Dentre vinte e um dos principais trabalhos científicos que recuperaram proteínas antigas de tecidos mineralizados fósseis (vertebrados, exceto Homo sapiens), publicados entre 2007 e 2022 (HUMPULA et al, 2007;CAPPELLINI et al, 2012;BUCKLEY, M, 2013;RYBCZYNSKI et al, 2013;BUCKLEY, M, 2015;HILL et al, 2015;WELKER et al, 2015;CLELAND et al, 2016;DEMARCHI et al, 2016;SCHROETER et al, 2017;WELKER et al, 2017;LAWLESS;RYBCZYNSKI, 2019;CAPPELLINI et al, 2019;PRESSLEE et al, 2019;WELKER et al, 2019WELKER et al, , 2020CAPPELLINI et al, 2020;COLLEARY et al, 2021;FROMENT et al, 2021;NOGUEIRA et al, 2021;CUCINA et al, 2022); apenas quatro o fizeram a partir de tecidos dentários (CAPPELLINI et al, 2019;WELKER et al, 2019WELKER et al, , 2020NOGUEIRA et al, 2021) e os demais a partir de tecido ósseo. Contudo, esse número vem crescendo nos últimos anos, devido à excelente preservação do esmalte no registro fóssil, e a possibilidade de inferências filogenéticas a partir das sequências de suas proteínas (DELGADO et al, 2008;CAPPELINI et al, 2018) Como já mencionado, a alta preservação em longos períodos de tempo, a composição física e molecular particular e a possibilidade de utilização de técnicas não-destrutivas (STEWART et al, 2016), fazem do esmalte um excelente tecido para recuperação de proteínas antigas (PORTO et al, 2011;STEWART et al, 2017;CAPPELINI et al, 2019;WELKER et al, 2019;NOGUEIRA et al, 2021).…”