Hominin phyloproteomics for the Pleistocene: PalaeoPROteomics of Skeletal Parts for Evolutionary Research

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Identifying the unidentified fauna enhances insights into hominin subsistence strategies during the Middle to Upper Palaeolithic transition

Publication . Sinet-Mathiot, Virginie; Rendu, William; Steele, Teresa E.; Spasov, Rosen; Madelaine, Stéphane; Renou, Sylvain; Soulier, Marie-Cécile; Martisius, Naomi L.; aldeias, vera; Endarova, Elena; Goldberg, Paul; McPherron, Shannon J. P.; Rezek, Zeljko; Sandgathe, Dennis; Sirakov, Nikolay; Sirakova, Svoboda; Soressi, Marie; Tsanova, Tsenka; Turq, Alain; Hublin, Jean-Jacques; Welker, Frido; Smith, Geoff M.

Understanding Palaeolithic hominin subsistence strategies requires the comprehensive taxonomic identification of faunal remains. The high fragmentation of Late Pleistocene faunal assemblages often prevents proper taxonomic identification based on bone morphology. It has been assumed that the morphologically unidentifiable component of the faunal assemblage would reflect the taxonomic abundances of the morphologically identified portion. In this study, we analyse three faunal datasets covering the Middle to Upper Palaeolithic transition (MUPT) at Bacho Kiro Cave (Bulgaria) and Les Cottes and La Ferrassie (France) with the application of collagen type I peptide mass fingerprinting (ZooMS). Our results emphasise that the fragmented component of Palaeolithic bone assemblages can differ significantly from the morphologically identifiable component. We obtain contrasting identification rates between taxa resulting in an overrepresentation of morphologically identified reindeer (Rangifer tarandus) and an underrepresentation of aurochs/bison (Bos/Bison) and horse/European ass (Equus) at Les Cottes and La Ferrassie. Together with an increase in the relative diversity of the faunal composition, these results have implications for the interpretation of subsistence strategies during a period of possible interaction between Neanderthals and Homo sapiens in Europe. Furthermore, shifts in faunal community composition and in carnivore activity suggest a change in the interaction between humans and carnivores across the MUPT and indicate a possible difference in site use between Neanderthals and Homo sapiens. The combined use of traditional and biomolecular methods allows (zoo)archaeologists to tackle some of the methodological limits commonly faced during the morphological assessment of Palaeolithic bone assemblages.

2023Journal article

Open access

Comparing extraction method efficiency for high-throughput palaeoproteomic bone species identification

Publication . Mylopotamitaki, Dorothea; Harking, Florian S.; Taurozzi, Alberto J.; Fagernäs, Zandra; Godinho, Ricardo Miguel; Smith, Geoff M.; Weiss, Marcel; Schüler, Tim; McPherron, Shannon P.; Meller, Harald; Cascalheira, João; Bicho, Nuno; Olsen, Jesper V.; Hublin, Jean-Jacques; Welker, Frido

High-throughput proteomic analysis of archaeological skeletal remains provides information about past fauna community compositions and species dispersals in time and space. Archaeological skeletal remains are a finite resource, however, and therefore it becomes relevant to optimize methods of skeletal proteome extraction. Ancient proteins in bone specimens can be highly degraded and consequently, extraction methods for well-preserved or modern bone might be unsuitable for the processing of highly degraded skeletal proteomes. In this study, we compared six proteomic extraction methods on Late Pleistocene remains with variable levels of proteome preservation. We tested the accuracy of species identification, protein sequence coverage, deamidation, and the number of post-translational modifications per method. We find striking differences in obtained proteome complexity and sequence coverage, highlighting that simple acid-insoluble proteome extraction methods perform better in highly degraded contexts. For well-preserved specimens, the approach using EDTA demineralization and protease-mix proteolysis yielded a higher number of identified peptides. The protocols presented here allowed protein extraction from ancient bone with a minimum number of working steps and equipment and yielded protein extracts within three working days. We expect further development along this route to benefit large-scale screening applications of relevance to archaeological and human evolution research.

2023Journal article

Open access