Deep-time phylogenetic inference by paleoproteomic analysis of dental enamel
Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › fagfællebedømt
In temperate and subtropical regions, ancient proteins are reported to survive up to about 2 million years, far beyond the known limits of ancient DNA preservation in the same areas. Accordingly, their amino acid sequences currently represent the only source of genetic information available to pursue phylogenetic inference involving species that went extinct too long ago to be amenable for ancient DNA analysis. Here we present a complete workflow, including sample preparation, mass spectrometric data acquisition and computational analysis, to recover and interpret million-year-old dental enamel protein sequences. During sample preparation, the proteolytic digestion step, usually an integral part of conventional bottom-up proteomics, is omitted to increase the recovery of the randomly degraded peptides spontaneously generated by extensive diagenetic hydrolysis of ancient proteins over geological time. Similarly, we describe other solutions we have adopted to (1) authenticate the endogenous origin of the protein traces we identify, (2) detect and validate amino acid variation in the ancient protein sequences and (3) attempt phylogenetic inference. Sample preparation and data acquisition can be completed in 3–4 working days, while subsequent data analysis usually takes 2–5 days. The workflow described requires basic expertise in ancient biomolecules analysis, mass spectrometry-based proteomics and molecular phylogeny. Finally, we describe the limits of this approach and its potential for the reconstruction of evolutionary relationships in paleontology and paleoanthropology.
Originalsprog | Engelsk |
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Tidsskrift | Nature Protocols |
Vol/bind | 19 |
Udgave nummer | 7 |
Sider (fra-til) | 2085-2116 |
ISSN | 1754-2189 |
DOI | |
Status | Udgivet - 2024 |
Bibliografisk note
Funding Information:
E.C., F.W. and J.R.-M. were supported by the VILLUM FONDEN (grant no. 17649). E.C., P.L.R. and J.V.O. were supported by the European Commission through the MSC European Training Network \u2018TEMPERA\u2019 (grant agreement no. 722606). E.C., I.P., C.K., R.S.P., P.P.M., F.S.H. and J.V.O. are supported by the European Commission through the MSC European Training Network \u2018PUSHH\u2019 (grant agreement no. 861389). E.C., A.J.T., M.M. and J.V.O. were supported by the European Research Council under the European Union\u2019s Horizon 2020 research and innovation program (grant agreement no. 101021361). F.W. has received funding from the European Research Council under the European Union\u2019s Horizon 2020 research and innovation program (grant agreement no. 948365). A.J.T. was supported by the Danish National Research Foundation award PROTEIOS (DNRF128). Work at The Novo Nordisk Foundation Center for Protein Research was funded in part by a generous donation from the Novo Nordisk Foundation (NNF14CC0001). We thank F. Racimo for critical reading of the manuscript and for the valuable comments and feedback.
Publisher Copyright:
© Springer Nature Limited 2024.
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