Deep-time phylogenetic inference by paleoproteomic analysis of dental enamel

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Pubblicato in:Nature Protocols vol. 19, no. 7 (Jul 2024), p. 2085
Autore principale: Taurozzi, Alberto J.
Altri autori: Rüther, Patrick L., Patramanis, Ioannis, Koenig, Claire, Sinclair Paterson, Ryan, Madupe, Palesa P., Harking, Florian Simon, Welker, Frido, Mackie, Meaghan, Ramos-Madrigal, Jazmín, Olsen, Jesper V., Cappellini, Enrico
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Nature Publishing Group
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024 7 |a 10.1038/s41596-024-00975-3  |2 doi 
035 |a 3076842482 
045 2 |b d20240701  |b d20240731 
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100 1 |a Taurozzi, Alberto J.  |u University of Copenhagen, Globe Institute, Copenhagen, Denmark (GRID:grid.5254.6) (ISNI:0000 0001 0674 042X) 
245 1 |a Deep-time phylogenetic inference by paleoproteomic analysis of dental enamel 
260 |b Nature Publishing Group  |c Jul 2024 
513 |a Journal Article 
520 3 |a 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.Key points<list list-type="bullet"><list-item></list-item>Paleoproteomics has shown that it is possible to obtain useful phylogenetic information from dental enamel proteins up to 2 million years old. They are heavily fragmented and chemically modified, making their recovery and analysis challenging.<list-item>The protocol describes how to (1) extract million-year-old dental enamel protein remains while minimizing contamination, (2) sequence them using high-resolution tandem mass spectrometry and (3) attempt otherwise so far impossible molecular-based phylogenetic inference.</list-item> 
653 |a Mass spectrometry 
653 |a Geological time 
653 |a Data acquisition 
653 |a Amino acids 
653 |a Genetic analysis 
653 |a Phylogenetics 
653 |a Dental enamel 
653 |a Phylogeny 
653 |a Proteomics 
653 |a Extinct species 
653 |a Amino acid sequence 
653 |a Workflow 
653 |a Paleontology 
653 |a Recovery 
653 |a Scientific imaging 
653 |a Nucleotide sequence 
653 |a Sample preparation 
653 |a Biomolecules 
653 |a Proteins 
653 |a Proteolysis 
653 |a Data processing 
653 |a Data analysis 
653 |a Inference 
653 |a Deoxyribonucleic acid--DNA 
653 |a Mass spectroscopy 
653 |a Peptides 
653 |a Environmental 
700 1 |a Rüther, Patrick L.  |u University of Copenhagen, Novo Nordisk Foundation Center for Protein Research, Copenhagen, Denmark (GRID:grid.5254.6) (ISNI:0000 0001 0674 042X) 
700 1 |a Patramanis, Ioannis  |u University of Copenhagen, Globe Institute, Copenhagen, Denmark (GRID:grid.5254.6) (ISNI:0000 0001 0674 042X) 
700 1 |a Koenig, Claire  |u University of Copenhagen, Novo Nordisk Foundation Center for Protein Research, Copenhagen, Denmark (GRID:grid.5254.6) (ISNI:0000 0001 0674 042X) 
700 1 |a Sinclair Paterson, Ryan  |u University of Copenhagen, Globe Institute, Copenhagen, Denmark (GRID:grid.5254.6) (ISNI:0000 0001 0674 042X) 
700 1 |a Madupe, Palesa P.  |u University of Copenhagen, Globe Institute, Copenhagen, Denmark (GRID:grid.5254.6) (ISNI:0000 0001 0674 042X) 
700 1 |a Harking, Florian Simon  |u University of Copenhagen, Novo Nordisk Foundation Center for Protein Research, Copenhagen, Denmark (GRID:grid.5254.6) (ISNI:0000 0001 0674 042X) 
700 1 |a Welker, Frido  |u University of Copenhagen, Globe Institute, Copenhagen, Denmark (GRID:grid.5254.6) (ISNI:0000 0001 0674 042X) 
700 1 |a Mackie, Meaghan  |u University of Copenhagen, Globe Institute, Copenhagen, Denmark (GRID:grid.5254.6) (ISNI:0000 0001 0674 042X); University of Copenhagen, Novo Nordisk Foundation Center for Protein Research, Copenhagen, Denmark (GRID:grid.5254.6) (ISNI:0000 0001 0674 042X) 
700 1 |a Ramos-Madrigal, Jazmín  |u University of Copenhagen, Globe Institute, Copenhagen, Denmark (GRID:grid.5254.6) (ISNI:0000 0001 0674 042X) 
700 1 |a Olsen, Jesper V.  |u University of Copenhagen, Novo Nordisk Foundation Center for Protein Research, Copenhagen, Denmark (GRID:grid.5254.6) (ISNI:0000 0001 0674 042X) 
700 1 |a Cappellini, Enrico  |u University of Copenhagen, Globe Institute, Copenhagen, Denmark (GRID:grid.5254.6) (ISNI:0000 0001 0674 042X) 
773 0 |t Nature Protocols  |g vol. 19, no. 7 (Jul 2024), p. 2085 
786 0 |d ProQuest  |t Health & Medical Collection 
856 4 1 |3 Citation/Abstract  |u https://www.proquest.com/docview/3076842482/abstract/embedded/H09TXR3UUZB2ISDL?source=fedsrch 
856 4 0 |3 Full Text - PDF  |u https://www.proquest.com/docview/3076842482/fulltextPDF/embedded/H09TXR3UUZB2ISDL?source=fedsrch