Palaeoproteomic investigation of an ancient human skeleton with abnormal deposition of dental calculus

Abstract

Detailed investigation of extremely severe pathological conditions in ancient human skeletons is important as it could shed light on the breadth of potential interactions between humans and disease etiologies in the past. Here, we applied palaeoproteomics to investigate an ancient human skeletal individual with severe oral pathology, focusing our research on bacterial pathogenic factors and host defense response. This female skeleton, from the Okhotsk period (i.e., fifth to thirteenth century) of Northern Japan, poses relevant amounts of abnormal dental calculus deposition and exhibits oral dysfunction due to severe periodontal disease. A shotgun mass-spectrometry analysis identified 81 human proteins and 15 bacterial proteins from the calculus of the subject. We identified two pathogenic or bioinvasive proteins originating from two of the three "red complex" bacteria, the core species associated with severe periodontal disease in modern humans, as well as two additional bioinvasive proteins of periodontal-associated bacteria. Moreover, we discovered defense response system-associated human proteins, although their proportion was mostly similar to those reported in ancient and modern human individuals with lower calculus deposition. These results suggest that the bacterial etiology was similar and the host defense response was not necessarily more intense in ancient individuals with significant amounts of abnormal dental calculus deposition.

Figure 1

(a) Map of Rebun Island. The map was drawn with the packages ggplot2 (version 3.4.0) and rnaturalearth (version 0.3.2) on version 4.2.2 of R. (b) Right buccal aspect of the HM2-HA-3 maxilla and mandible. A red arrow indicates the sampled calculus (i.e., from the lower right permanent first incisor).

Figure 2

Carbon and nitrogen stable isotopic results of faunal and human bone collagen.

Figure 3

Venn diagrams of (a) human proteins and (b) bacterial taxa identified in the ancient dental calculus of HM2-HA-3 (this study) as well as in the dental calculus samples from medieval Dalheim and modern patients.

Figure 4

Results of PANTHER (a) biological process and (b) protein class analysis of protein groups identified in the dental calculus of HM2-HA-3 (this study) as well as in the dental calculus samples from medieval Dalheim and modern patients.