Sexing remains of a Byzantine ascetic burial using enamel proteomics

The identification of sex in human remains is often met with challenges due to poor preservation of the remains. In this research we address the question of sex determination using proteome analysis of tooth enamel of a Byzantine burial that exhibited the practice of asceticism (bodily punishment). This practice has generally been associated with males, an issue we wanted to confirm. We applied peptidomic analysis of tooth enamel to determine the sample’s sex and confirmed our results by Label Free Quantification (LFQ). We analyzed also AmelX hydrolysis patterns. The absence of an AmelY unique spectra enabled us to classify the remains as very probably those of a female. Our findings indicate that the ancient AmelX exhibited non-enzymatic hydrolysis, which is also reflected in significantly shorter peptides. Additionally, we look further into the specificity actual places of cleavage. Our research shows that a simple peptidomics workflow is capable of determining sex and provides a robust solution for the analysis of samples from contexts suffering from poor bone preservation. The presence of a female ascetic introduces the necessity for additional studies of this phenomenon and its sociological implications. • Identification of Byzantine Female Ascetic : Dental proteomics suggest female sex by the absence of male-specific peptides. • First Documentation of Female Self-Torment Rituals : Unveils women’s role in extreme ascetic practices. • Significance for Historical and Sociological Research : Highlights gender dynamics and religious rituals in Byzantine society.