- Palaeoproteomics & Bioarchaeology, Department of Life Sciences and Systems Biology, University of Turin, Turin, Italy
- Archaeometry, Bioarchaeology, Dating, Raw materials, Zooarchaeology
Parchment Glutamine Index (PQI): A novel method to estimate glutamine deamidation levels in parchment collagen obtained from low-quality MALDI-TOF data
Assessing glutamine deamination in ancient parchment samplesRecommended by Beatrice Demarchi based on reviews by Maria Codlin and 3 anonymous reviewers
Data authenticity and approaches to data authentication are crucial issues in ancient protein research. The advent of modern mass spectrometry has enabled the detection of traces of ancient biomolecules contained in fossils, including protein sequences. However, detecting proteins in ancient samples does not equate to demonstrating their endogenous nature: instead, if the mechanisms that drive protein preservation and degradation are understood, then the extent of protein diagenesis can be used for evaluating preservational quality, which in turn may be related to the authenticity of the protein data.
The post-mortem deamidation of asparaginyl and glutamyl residues is a key degradation reaction, which can be assessed effectively on the basis of mass spectrometry data, and which has accrued a long history of research, both in terms of describing the mechanisms governing the reactions and with regard to the best strategies for assessing and quantifying the extent of glutamine (Gln) and asparagine (Asn) deamidation in ancient samples (Pal Chowdhury et al., 2019; Ramsøe et al., 2021, 2020; Schroeter and Cleland, 2016; Simpson et al., 2016; Solazzo et al., 2014; Welker et al., 2016; Wilson et al., 2012).
In their paper, Nair and colleagues (2022) build on this wealth of knowledge and present a tool for quantifying the extent of Gln deamidation in parchment. Parchment is a collagen-based material which can yield extraordinary insights into manuscript manufacturing practices in the past, as well as on the daily lives of the people who assembled and used them (“biocodicology”) (Fiddyment et al., 2021, 2019, 2015; Teasdale et al., 2017). Importantly, the extent of deamidation can be directly related to the quality of the parchment produced: rapid direct deamidation of Gln is induced by the liming process, therefore high extents of deamidation are linked to prolonged exposure to the high pH conditions which are typical of liming, thus implying lower-quality parchment.
Nair et al.’s approach focuses on collagen peptides which are typically detected during MALDI-TOF mass spectrometry analyses of parchment and build a simple three-step workflow able to yield an overall index of deamidation for a sample (the parchment glutamine index - PQI) 一 taking into account that different Gln residues degrade at different rates according to their micro-chemical environment. The first step involves pre-processing the MALDI spectra, since Nair et al. are specifically interested in maximising information which can be obtained by low-quality data. The second step builds on well-established methods for quantifying Q → E from MALDI-TOF data by modelling the convoluted isotope distributions (Wilson et al., 2012). Once relative rates of deamidation in selected peptides within a given sample are calculated, the third step uses a mixed effects model to combine the individual deamidation estimates and to obtain an overall estimate of the deamidation for a parchment sample (PQI).
The PQI can be used effectively for assessing parchment quality, as the authors show for the dataset from Orval Abbey. However, PQI could also have wider applications to the study of processed collagen, which is widely used in the food and pharmaceutical industries. In general, the study by Nair et al. is a welcome addition to a growing body of research on protein diagenesis, which will ultimately improve models for the assessment of the authenticity of biomolecular data in archaeology.
Chowdhury, P.M., Wogelius, R., Manning, P.L., Metz, L., Slimak, L., and Buckley, M. 2019. Collagen deamidation in archaeological bone as an assessment for relative decay rates. Archaeometry 61:1382–1398. https://doi.org/10.1111/arcm.12492
Fiddyment, S., Goodison, N.J., Brenner, E., Signorello, S., Price, K., and Collins, M.J.. 2021. Girding the loins? Direct evidence of the use of a medieval parchment birthing girdle from biomolecular analysis. bioRxiv. https://doi.org/10.1098/rsos.202055
Fiddyment,S., Holsinger, B., Ruzzier, C., Devine, A., Binois, A., Albarella, U., Fischer, R., Nichols, E., Curtis, A., Cheese, E., Teasdale, M.D., Checkley-Scott, C., Milner, S.J., Rudy, K.M., Johnson, E.J., Vnouček, J., Garrison, M., McGrory, S., Bradley, D.G., and Collins, M.J. 2015. Animal origin of 13th-century uterine vellum revealed using noninvasive peptide fingerprinting. Proc Natl Acad Sci U S A 112:15066–15071. https://doi.org/10.1073/pnas.1512264112
Fiddyment, S., Teasdale, M.D., Vnouček, J., Lévêque, É., Binois, A., and Collins, M.J. 2019. So you want to do biocodicology? A field guide to the biological analysis of parchment. Heritage Science 7:35. https://doi.org/10.1186/s40494-019-0278-6
Nair, B., Rodríguez Palomo, I., Markussen, B., Wiuf, C., Fiddyment, S., and Collins, M. Parchment Glutamine Index (PQI): A novel method to estimate glutamine deamidation levels in parchment collagen obtained from low-quality MALDI-TOF data. BiorRxiv, 2022.03.13.483627, ver. 6 peer-reviewed and recommended by Peer community in Archaeology. https://doi.org/10.1101/2022.03.13.483627
Ramsøe, A., Crispin, M., Mackie, M., McGrath, K., Fischer, R., Demarchi, B., Collins, M.J., Hendy, J., and Speller, C. 2021. Assessing the degradation of ancient milk proteins through site-specific deamidation patterns. Sci Rep 11:7795. https://doi.org/10.1038/s41598-021-87125-x
Ramsøe, A., van Heekeren, V., Ponce, P., Fischer, R., Barnes, I., Speller, C., and Collins, M.J. 2020. DeamiDATE 1.0: Site-specific deamidation as a tool to assess authenticity of members of ancient proteomes. J Archaeol Sci 115:105080. https://doi.org/10.1016/j.jas.2020.105080
Schroeter, E.R., and Cleland, T.P. 2016. Glutamine deamidation: an indicator of antiquity, or preservational quality? Rapid Commun Mass Spectrom 30:251–255. https://doi.org/10.1002/rcm.7445
Simpson, J.P., Penkman, K.E.H., and Demarchi, B. 2016. The effects of demineralisation and sampling point variability on the measurement of glutamine deamidation in type I collagen extracted from bone. J Archaeol Sci 69: 29-38. https://doi.org/10.1016/j.jas.2016.02.002
Solazzo, C., Wilson, J., Dyer, J.M., Clerens, S., Plowman, J.E., von Holstein, I., Walton Rogers, P., Peacock, E.E., and Collins, M.J. 2014. Modeling deamidation in sheep α-keratin peptides and application to archeological wool textiles. Anal Chem 86:567–575. https://doi.org/10.1021/ac4026362
Teasdale, M.D., Fiddyment, S., Vnouček, J., Mattiangeli, V., Speller, C., Binois, A., Carver, M., Dand, C., Newfield, T.P., Webb, C.C., Bradley, D.G., and Collins M.J. 2017. The York Gospels: a 1000-year biological palimpsest. R Soc Open Sci 4:170988. https://doi.org/10.1098/rsos.170988
Welker, F., Soressi, M.A., Roussel, M., van Riemsdijk, I., Hublin, J.-J., and Collins, M.J. 2016. Variations in glutamine deamidation for a Châtelperronian bone assemblage as measured by peptide mass fingerprinting of collagen. STAR: Science & Technology of Archaeological Research 3:15–27. https://doi.org/10.1080/20548923.2016.1258825
Wilson, J., van Doorn, N.L., and Collins, M.J. 2012. Assessing the extent of bone degradation using glutamine deamidation in collagen. Anal Chem 84:9041–9048. https://doi.org/10.1021/ac301333t
The management of symbolic raw materials in the Late Upper Paleolithic of South-Western France: a shell ornaments perspective
Caching up with the study of the procurement of symbolic raw materials in the Upper PalaeolithicRecommended by Beatrice Demarchi based on reviews by Begoña Soler Mayor , Catherine Dupont and Lawrence Straus
The manuscript "The management of symbolic raw materials in the Late Upper Paleolithic of South-Western France: a shell ornaments perspective" by Solange Rigaud and colleagues (Rigaud et al. 2022) is a perfect demonstration that appropriate scientific methodologies can be used effectively in order to enhance the historical value of findings from “old” collections, despite the lack of secure stratigraphic and contextual data. The shell assemblage (n = 377) investigated here (from Rochereil, Dordogne) had been excavated during the first half of the 20th century (Jude 1960) and reported in 1993 (Taborin 1993), but only this recent analysis revealed that it was composed of largely unmodified mollusc shells, most of allochthonous origin. Rigaud et al. interpret this finding as the raw materials used to produce personal ornaments. This is especially significant, because the focus of research has been on the manufacture, use and exchange of personal ornaments in prehistory, much less so on the procurement of the raw materials. As such, the manuscript adds substantially to the growing literature on Magdalenian social networks.
The authors carried out detailed taxonomic analysis based on morphological and morphometric characteristics and identified at least nine different species, including Dentalium sp., Ocenebra erinaceus, Tritia reticulata and T. gibbosula, as well as some bivalve specimens (Mytilus, Glycymeris, Spondylus, Pecten). Most of the species are commonly found in personal ornament assemblages from the Magdalenian, reflecting intentional selection (also shown by the size sorting of some of the taxa), and cultural continuity. However, microscopic examinations revealed securely-identified anthropogenic modifications on a very limited number of specimens: one Glycymeris valve (used as an ochre container), one Cardiidae valve (presence of a groove), one perforated Tritia gibbosula and two perforated Tritia reticulata bearing striations. The authors interpret this combination of anthropogenic vs natural “signals” as signifying that the assemblage represents raw material selected and stored for further processing.
Assessing the provenance and age of the shells is therefore paramount: the shells found at Rochereil belong to species that can be found on both the Atlantic and Mediterranean coasts. Assuming that molluscan taxa distribution in the past is comparable to that for the present day, this implies the exploitation of two catchment areas and long-distance transportation to the site: taking sea-level changes into account, during the Magdalenian the Mediterranean used to lie at a distance of 350 km from Rochereil, and the Atlantic was not significantly closer (~200 km). Importantly, exploitation of fossil shells cannot be discounted on the basis of the data presented here; direct dating of some of the specimens (e.g. by radiocarbon, or amino acid racemisation geochronology) would be beneficial to clarify this issue and in general to improve chronological control on the accumulation of shells. Nonetheless, the authors argue that the closest fossil deposits also lie more than 200 km away from the site, thus the material is allochthonous in origin.
In synthesis, the Rochereil assemblage represents an important step towards a better understanding of the procurement chain and of the production of ornaments during the European Upper Palaeolithic.
Jude, P. E. (1960). La grotte de Rocherreil: station magdalénienne et azilienne, Masson.
Rigaud, S., O'Hara, J., Charles, L., Man-Estier, E. and Paillet, P. (2022) The management of symbolic raw materials in the Late Upper Paleolithic of South-Western France: a shell ornaments perspective. SocArXiv, z7pqg, ver. 4 peer-reviewed and recommended by Peer community in Archaeology. https://doi.org/10.31235/osf.io/z7pqg
Taborin, Y. (1993). La parure en coquillage au Paléolithique, CNRS éditions.
A way to break bones? The weight of intuitiveness
Breaking bones: Nature or Culture?Recommended by Beatrice Demarchi and Reuven Yeshurun based on reviews by Terry O'Connor, Alan Outram and 1 anonymous reviewer
The nature of breaking long bones for obtaining marrow is important in Paleolithic archaeology, due to its widespread, almost universal, character. Provided that hammer-stone percussion marks can be correctly identified using experimental datasets (e.g., ), the anatomical location and count of the marks may be taken to reflect recurrent “cultural” traditions in the Paleolithic . Were MP humans breaking bones intuitively or did they abide by a strict “protocol”, and, if the latter, was this protocol optimized for marrow retrieval or geared towards another, less obvious goal?
The new preprint by Vettese and colleagues  took a novel approach to this question, by conducting an experiment which involved novice butchers, relying on nothing but their intuition, and recording the way in which they broke marrow-rich bones. Some variability was noted in the “intuitive” patterns; indeed, future studies replicating this experiment and adding more variables such as more experienced butchers and non-bovid bones are warranted. Similarities in the means by which novice butchers break bones was also observed, and especially telling is the strong effect of anatomical features in most bones, except for the femur.
This paper provides a baseline for location analyses of percussion marks. Their dataset may therefore be regarded as a null hypothesis according to which the archaeological data could be tested. If Paleolithic patterns of percussion marks differ from Vettese et al.’s  “intuitive” patterns, then the null hypothesis is disproved and one can argue in favor of a learned pattern. The latter can be a result of ”culture”, as Vettese et al.  phrase it, in the sense of nonrandom action that draws on transmitted knowledge. Such comparisons bear a great potential for understanding the degree of technological behavior in the Paleolithic by factoring out the “natural” constraints of bone breakage patterns. Vettese et al. [3: fig. 14] started this discourse by comparing their experimental dataset to some Middle and Upper Paleolithic faunas; we are confident that many other studies will follow.
Pickering, T.R., Egeland, C.P., 2006. Experimental patterns of hammerstone percussion damage on bones: Implications for inferences of carcass processing by humans. J. Archaeol. Sci. 33, 459–469. https://doi.org/10.1016/j.jas.2005.09.001
Blasco, R., Rosell, J., Domínguez-Rodrigo, M., Lozano, S., Pastó, I., Riba, D., Vaquero, M., Peris, J.F., Arsuaga, J.L., de Castro, J.M.B., Carbonell, E., 2013. Learning by Heart: Cultural Patterns in the Faunal Processing Sequence during the Middle Pleistocene. PLoS One 8, e55863. https://doi.org/10.1371/journal.pone.0055863
Vettese, D., Stavrova, T., Borel, A., Marin, J., Moncel, M.-H., Arzarello, M., Daujeard, C. (2020) A way to break bones? The weight of intuitiveness. BioRxiv, 011320, ver. 4 peer-reviewed and recommended by PCI Archaeology. https://doi.org/10.1101/2020.03.31.011320