Biology subjects

Welker, F.

Publications and source records attributed to Welker, F..

13 recordsLinked to original sources

bioRxiv2026

No signal of a super-archaic origin of the Denisovan AMBN gene, a comment on the protein affinity of Homo erectus and Denisovan enamel proteins.

Homo erectus is a species that occupies a central role in the study of human evolution. To investigate its taxonomic identity, Fu et al. 2026 extracted and sequenced enamel proteins from 6 fossils assigned to Homo erectus, originating from 3 localities in China and dated to around 400 thousand years ago. Unexpectedly, all samples possess an amino acid variant on the enamel protein ameloblastin (AMBN) which is uniquely shared with more recent Denisovan fossils and a subset of present day humans who are known to carry introgressed Denisovan ancestry. The samples also showcase an additional, novel variant on the same enamel protein. To explain this result, Fu et al. 2026 propose a model where the sampled Homo erectus population (or its recent ancestors) interbred with later arriving Denisovans, introducing one of the two AMBN variants into the Denisovan population. While this model fits our overall understanding of the interactions between these archaic populations, it rests on the assumption that the Denisovan AMBN variant has an archaic, Homo erectus-like, source. Here we show that the AMBN gene of late Denisovans has no signal of introgression from a super-archaic source, making the suggested model unlikely for this gene. We also show that the AMBN gene of Denisova 25, an earlier Denisova, does show a signal of introgression, but one that better matches Neanderthals and modern humans, rather than a super-archaic source. We propose a number of alternative models that could help explain the observed affinity between the sampled H. erectus and Denisovans without requiring the introgression of AMBN from an archaic source into Denisovans. We discuss their strengths and weaknesses and how new data could help resolve them.

evolutionary biology↗

bioRxiv2026

Archaeological preservation of amelogenesis pathways

Dental enamel, the hardest mineralised tissue in the human body, has proven to be an excellent source of ancient proteins, which have been found to survive within dental enamel for at least twenty million years. In archaeological and palaeontological contexts, the enamel proteome is generally considered to be rather small, consisting of about twelve proteins, most of which are unique to enamel. During amelogenesis these proteins undergo in vivo digestion by matrix metalloproteinase 20 (MMP20) and kallikrein 4 (KLK4) as well as serine phosphorylation by family with sequence similarity member 20-C (FAM20C) that alter their characteristics. Gaining knowledge of the previously understudied influence of amelogenesis on the archaeological human dental enamel proteome could benefit various palaeoproteomic analysis, especially in an human evolutionary context. Here we present archaeological dental enamel proteomes and explore protein cleavage patterns and sequence coverage to estimate the effects of in vivo digestion, as well as explore phosphorylation patterns. Additionally, we present a new marker based on phosphorylation to estimate genetic sex.

evolutionary biology↗

bioRxiv2026

Beyond dairy: Identification of dental enamel proteins in ancient human dental calculus

Ancient human dental calculus is one of the richest archives of archaeological biomolecular information, providing direct evidence of diet, oral health, and the oral microbiome. Proteomic analyses of this biological matrix have so far focused mainly on oral microbes and dietary proteins, with milk proteins such as beta-lactoglobulin (BLG) providing the largest corpus of proteomic evidence. Despite the close relation between the various stages of dental calculus formation and mineralization with the dental enamel surface, proteins from the dental enamel matrix have not previously been reported outside of dental enamel tissue. Here we reanalysed 498 ancient dental calculus proteomes from 14 published studies (n=434 individuals) reporting the presence of BLG, spanning from the Neolithic to the Victorian Era and applying different protein extraction protocols (FASP, GASP, SP3 and in-solution digestion). Dental enamel matrix proteins were identified in ten studies (n=37 individuals), with amelogenin being the most frequently detected. Enamel peptides occurred more often in studies that applied SP3, although amelogenin was successfully identified through both SP3 and FASP. Structural proteins, including enamelin, ameloblastin, and MMP20, were also identified. The detection of AMELX and AMELY peptide sequences provided new insights into cases where the sex was previously undetermined. These findings establish dental enamel proteins as a new category of biomolecules detected in dental calculus, broadening its application beyond diet and microbiome studies to possible sex estimation. HighlightsO_LIDental calculus entraps oral microbes along with endogenous and exogenous particles during formation and mineralization C_LIO_LIWe conduct reanalysis of 14 published ancient dental calculus studies (n = 434 individuals) spanning the Neolithic to Victorian Era C_LIO_LIDental enamel proteins AMELX, AMELY, AMBN, COL17A1, ENAM and MMP20 are identified in ancient human dental calculus C_LIO_LIAmelogenin was the most frequently detected enamel protein C_LIO_LIWe expand dental calculus palaeoproteomics beyond diet and oral microbiome to potentially include sex estimation C_LI

evolutionary biology↗

bioRxiv2025

Composition variations in archaeological human bone proteomes

Sampling strategies within the field of skeletal palaeoproteomics are often based on specimen availability. Knowledge of bone biology might assist in improving sample selection strategies and minimise unnecessary sampling of precious (hominin) material. We study ten bone sample locations across four bone elements, for a total of 10 adult, archaeological human skeletons. We compare bone proteome composition and modification for skeletal elements formed through endochondral and intramembranous ossification, as well as cortical-trabecular bone pairs of three skeletal locations. We observe minimal differences in bones formed through the two ossification processes, outside of the exclusive presence of cartilage-related proteins in endochondral bone samples. We observe higher protein concentrations, a larger number of protein groups and peptides, and lower rates of deamidation in cortical bone compared to trabecular bone proteomes, this indicates that cortical bone provides a better preservation environment compared to trabecular bone. Throughout our analysis, the petrous bone stands out, with the largest and most complex proteomes recovered for all studied individuals. Formed through endochondral ossification, the petrous bone undergoes minimal turnover during life. Our observations indicate that the petrous bone is the ideal source of ancient protein sequence information.

evolutionary biology↗

bioRxiv2025

Identification and removal of contamination in palaeoproteomic analysis of dental enamel

Analysis of archaeological and palaeontological dental enamel allows for the study of taxonomy, phylogenetic relationships, and genetic sex of Pleistocene fauna and hominin remains. However, high-quality data is required for the study of degraded, chemically modified ancient proteomes. Protein contamination may hinder acquisition of such data, due to the abundance and better preservation of modern contaminating proteins. Here, we artificially contaminate a Pleistocene woolly rhinoceros tooth in order to investigate decontamination of archaeological dental enamel. We find that although the contaminating proteins are not identified, likely due to a lack of denaturation and digestion steps in archaeological dental enamel protein extraction protocols, contamination leads to significant loss of endogenous proteomic information. We thereafter compare five published decontamination methods, and find that a simple water or bleach wash prior to demineralization is most efficient at removing protein contamination. The water wash does not affect the endogenous proteome, while a bleach wash may lead to a loss of shorter intercrystalline peptides. We therefore conclude that decontamination is necessary in the study of ancient dental enamel, to ensure maximal retrieval of endogenous proteomic information, and can easily be achieved by incorporating a water or bleach wash step into the protein extraction protocol.

evolutionary biology↗

bioRxiv2025

New methods on the block: Taxonomic identification of archaeological bones in resin-embedded sediments through palaeoproteomics

The integration of biomolecular studies of past organisms with geoarchaeological studies can significantly improve our understanding of the relative chronology and context of archaeologically (in)visible behaviours. However, the complexity and sedimentological heterogeneity of archaeological deposits at a microscopic scale is often not taken into consideration in biomolecular studies. Here, we investigate the preservation and retrieval of palaeoproteomic data from bone fragments embedded in Pleistocene resin-impregnated sediment blocks. We show that resin impregnation has minimal effect on skeletal protein taxonomic identifications in modern skeletal material, but observe an increase in oxidation-related post-translational modifications. We then successfully retrieve proteins from resin-impregnated blocks from the Palaeolithic sites of Bacho Kiro Cave, La Ferrassie and Quincay. The taxonomic identifications of minute bones encased in resin are in line with previous analyses of the faunal communities of these sites, with a diversity of taxa (Bos sp./Bison sp., Equus sp., Ursus sp., and Caprinae) observed at a microscale in Bacho Kiro. This differs from results from La Ferrassie where most of the samples are identified as a single taxon (Bos sp./Bison sp.) across different areas of the site. The block from Quincay only provided taxonomic identification of two out of eleven bone-derived samples, likely due to diagenesis. Our work indicates that palaeoproteomes can be retrieved from bone fragments at a microstratigraphic resolution, enabling the detailed study of faunal community composition at a scale that more closely matches that of past human occupations. Significance StatementResin-embedded sediment blocks are widely used in archaeology and soil sciences to reconstruct past environments and human behavior, but their potential for biomolecular analysis is underexplored. Here, we demonstrate that ancient proteins can be successfully retrieved from bone fragments embedded in resin-impregnated sediment blocks from Pleistocene archaeological sites. Our findings show that resin impregnation has a minimal impact on protein recovery and that palaeoproteomics enables taxonomic identification of Pleistocene bone fragments at a microstratigraphic scale. This approach allows for reconstructing past faunal communities with unprecedented detail, improving our understanding of ancient ecosystems and the environmental contexts of early hominin occupations.

evolutionary biology↗

bioRxiv2024

Ancient biomolecular analysis of 39 mammoth individuals from Kostenki 11-Ia elucidates Upper Palaeolithic human resource use

Circular structures made from woolly mammoth bones are found across Ukraine and west Russia, yet the origin of the bones remains uncertain. We present ten new mammoth radiocarbon dates from the largest circular structure at Kostenki 11-Ia, identifying two mammoth mandibles [~]200-1,200 years older than the other dated materials from the site, suggesting skeletal material from long-dead individuals was scavenged and used in the site construction. Biomolecular sexing of 30 individuals showed a predominance of females, suggesting the Kostenki mammoths are primarily from herds. We identify six mitochondrial lineages across 16 samples, showing they are not all from the same matriline. Integrating biomolecular sexing with stable{delta} 13C and{delta} 15N isotope analysis, we find no isotopically-differentiated resource use by females and males, providing the first analysis of foraging differences between sexes in any Late Pleistocene megafauna. Our study highlights the significance of integrating ancient biomolecular approaches in archaeological inference. TeaserIntegrating 14C dating, ancient DNA, palaeoproteomics, and stable isotopes improves our understanding of Kostenki 11-Ia

evolutionary biology↗

bioRxiv2024

Cleaning the Dead: Optimized decontamination enhances palaeoproteomic analyses of a Pleistocene hominin tooth from Khudji, Tajikistan

The study of ancient proteins preserved in a range of archaeological, cultural heritage, and palaeontological materials is increasingly contributing to our understanding of human evolution and archaeological research questions. Many of the specimens studied have been excavated and stored for a significant duration prior to their proteomic analysis. Human handling and storage environments therefore provide ample opportunities for protein contamination onto and into specimens of interest to palaeoproteomic studies. As such, modern protein contamination limits access to endogenous proteomes. Here, we compare five approaches of bone protein decontamination applied to a Pleistocene Equus sp. bone fragment contaminated with a modern dog salivary proteome. We find that all tested methods reduce the protein contamination, but with different efficiencies. We find that a brief bleach wash is the most effective approach in removing modern protein contamination, and that no additional damage is caused to the endogenous proteome by this treatment. Next, we apply this approach to a hominin tooth found at Khudji, a Late Pleistocene archaeological site in Tajikistan. We demonstrate that a brief bleach wash removes almost all human skin protein contamination while retaining the endogenous hominin dentine proteome. Subsequent phylogenetic analysis of the Khudji dentine proteome allowed determination that the specimen is likely not a Denisovan, but still leaves ambiguity between an assignment to either modern humans or Neanderthals.

evolutionary biology↗

bioRxiv2024

Palaeoproteomic identification of a whale bone tool from Bronze Age Heiloo, the Netherlands

Identification of the taxonomic origin of bone tools is an important, but often complicated, component of studying past societies. The species used for bone tool production provide insight into what species were exploited, potentially how, and for what purpose. Additionally, the choice of species may have important implications for the place of the tool within the larger toolkit. However, the taxonomic identification of bone tools is often unsuccessful based on morphology. Here we apply three palaeoproteomic techniques, ZooMS, SPIN-like data analysis and a targeted database search to narrow down the taxonomic identification of an unusually large Bronze Age bone tool from Heiloo, the Netherlands, to the North Atlantic right whale (Eubalaena glacialis). Additionally, the tool was investigated for use-wear, which showed that it was likely used for the processing of plant fibres. The assignment of the tool as whale bone adds support to the exploitation of whales by coastal Bronze Age populations, not just for meat, as previously suggested, but also for bone as a resource for tool production. We know of no other parallel of a bone tool such as this in terms of size, use, hafting, and taxonomic identity.

evolutionary biology↗

bioRxiv2023

Enamel proteins reveal biological sex and genetic variability within southern African Paranthropus

The evolutionary relationships among extinct African hominin taxa are highly debated and largely unresolved, due in part to a lack of molecular data. Even within taxa, it is not always clear, based on morphology alone, whether ranges of variation are due to sexual dimorphism versus potentially undescribed taxonomic diversity. For Paranthropus robustus, a Pleistocene hominin found only in South Africa, both phylogenetic relationships to other taxa 1,2 and the nature of intraspecific variation 3-6 are still disputed. Here we report the mass spectrometric (MS) sequencing of enamel proteomes from four ca. 2 million year (Ma) old dental specimens attributed morphologically to P. robustus, from the site of Swartkrans. The identification of AMELY-specific peptides and semi-quantitative MS data analysis enabled us to determine the biological sex of all the specimens. Our combined molecular and morphometric data also provide compelling evidence of a significant degree of variation within southern African Paranthropus, as previously suggested based on morphology alone 6. Finally, the molecular data also confirm the taxonomic placement of Paranthropus within the hominin clade. This study demonstrates the feasibility of recovering informative Early Pleistocene hominin enamel proteins from Africa. Crucially, it also shows how the analysis of these proteins can contribute to understanding whether hominin morphological variation is due to sexual dimorphism or to taxonomic differences. We anticipate that this approach can be widely applied to geologically-comparable sites within South Africa, and possibly more broadly across the continent.

evolutionary biology↗

bioRxiv2021

Spectrum graph-based de-novo sequencing algorithm MaxNovo achieves high peptide identification rates in collisional dissociation MS/MS spectra

We describe MaxNovo, a novel spectrum graph-based peptide de-novo sequencing algorithm integrated into the MaxQuant software. It identifies complete sequences of peptides as well as sequence tags that are incomplete at one or both of the peptide termini. MaxNovo searches for the highest-scoring path in a directed acyclic graph representing the MS/MS spectrum with peaks as nodes and edges as potential sequence constituents consisting of single amino acids or pairs. The raw score is a sum of node and edge weights, plus several reward scores, for instance, for complementary ions or protease compatibility. For search-engine identified peptides, it correlates well with the Andromeda search engine score. We use a particular score normalization and the score difference between the first and second-best solution to define a combined score that integrates all available information. To evaluate its performance, we use a human cell line dataset and take as ground truth all Andromeda-identified MS/MS spectra with an Andromeda score of at least 100. MaxNovo outperforms other software in particular in the high-sensitivity range of precision-coverage plots. We also identify incomplete sequence tags and study their statistical properties. Next, we apply MaxNovo to ion mobility-coupled time of flight data. Here we achieve excellent performance as well, except for potential swaps of the two amino acids closest to the C-terminus, which are not well resolved due to the low end of the mass range in MS/MS spectra in this dataset. We demonstrate the applicability of MaxNovo to palaeoproteomics samples with a Late Pleistocene hominin proteome dataset that was generated using three proteases. Interestingly, we did not use any machine learning in the construction of MaxNovo, but implemented expert domain knowledge directly in the definition of the score. Yet, it performs as good as or better than the leading deep learning-based algorithm.

bioinformatics↗

bioRxiv2021

SPIN - Species by Proteome INvestigation

Genetic species determination has become an indispensable tool in forensics, archaeology, ecology, and food authentication. The available methods are either suited for detecting a single taxon across many samples or for screening a wide range of species across a few samples. Here, we introduce "Species by Proteome INvestigation" (SPIN), a proteomics workflow capable of querying over 150 mammalian species in 7.2 minutes of mass spectrometry (MS) analysis. Streamlined and automated sample preparation by protein aggregation capture, high-speed chromatography and data-independent acquisition, and a confident species inference algorithm facilitate processing hundreds of samples per day. We demonstrate the correct classification of known references, reproducible species identification in degraded Iron-Age material from Scandinavia, and test the limits of our methods with Middle and Upper Palaeolithic bones from Southern European sites with late Neanderthal occupation. While this initial study is focused on modern and archaeological mammalian bone, SPIN will be open and expandable with other biological tissues and taxa.

biochemistry↗

bioRxiv2018

Early Pleistocene enamel proteome sequences from Dmanisi resolve Stephanorhinus phylogeny.

Ancient DNA (aDNA) sequencing has enabled unprecedented reconstruction of speciation, migration, and admixture events for extinct taxa1. Outside the permafrost, however, irreversible aDNA post-mortem degradation2 has so far limited aDNA recovery within the {small tilde}0.5 million years (Ma) time range3. Tandem mass spectrometry (MS)-based collagen type I (COL1) sequencing provides direct access to older biomolecular information4, though with limited phylogenetic use. In the absence of molecular evidence, the speciation of several Early and Middle Pleistocene extinct species remain contentious. In this study, we address the phylogenetic relationships of the Eurasian Pleistocene Rhinocerotidae5-7 using {small tilde}1.77 million years (Ma) old dental enamel proteome sequences of a Stephanorhinus specimen from the Dmanisi archaeological site in Georgia (South Caucasus)8. Molecular phylogenetic analyses place the Dmanisi Stephanorhinus as a sister group to the woolly (Coelodonta antiquitatis) and Mercks rhinoceros (S. kirchbergensis) clade. We show that Coelodonta evolved from an early Stephanorhinus lineage and that this genus includes at least two distinct evolutionary lines. As such, the genus Stephanorhinus is currently paraphyletic and its systematic revision is therefore needed. We demonstrate that Early Pleistocene dental enamel proteome sequencing overcomes the limits of ancient collagen- and aDNA-based phylogenetic inference, and also provides additional information about the sex and taxonomic assignment of the specimens analysed. Dental enamel, the hardest tissue in vertebrates, is highly abundant in the fossil record. Our findings reveal that palaeoproteomic investigation of this material can push biomolecular investigation further back into the Early Pleistocene.

evolutionary biology↗