Abstract
Recent work has disclosed the critical role played by enamel peptides in sex classification of old skeletal remains. In particular, protein AMELY (amelogenin isoform Y) is present in the enamel dental tissue of male individuals only, while AMELX (isoform X) can be found in both sexes. AMELY can be easily detected by LC-MS/MS in the ion extracted chromatograms of the SM(ox)IRPPY peptide (monoisotopic [M + 2 H]+2 mass = 440.2233 m/z). In this paper, we exploited the dimorphic features of the amelogenin protein to determine the sex of the so-called ‘Lovers of Modena’, two Late Antique individuals whose skeletons were intentionally buried hand-in-hand. Upon discovery, mass media had immediately assumed they were a male-female couple, even if bad preservation of the bones did not allow an effective sex classification. We were able to extract proteins from the dental enamel of both individuals (~1600 years old) and to confidently classify them as males. Results were compared to 14 modern and archaeological control samples, confirming the reliability of the ion chromatogram method for sex determination. Although we currently have no information on the actual relationship between the ‘Lovers of Modena’ (affective? Kin-based?), the discovery of two adult males intentionally buried hand-in-hand may have profound implications for our understanding of funerary practices in Late Antique Italy.
Highlights
A paper by Stewart et al.[7] has revolutionized the way to achieve sex determination of skeletons from archaeological and forensic contexts, thanks to the enamel proteins. One of these proteins is amelogenin, whose gene is translated into two isoforms linked to sexual chromosomes[8], namely AMELX and AMELY, which constitute 90% of the entire enamel proteome[9]
By liquid-chromatography mass spectrometry (LC-MS/MS), a relatively rapid and cost-effective technique, it is possible to check the occurrence of AMELX and/or AMELY isoforms within enamel samples in order to rapidly estimate the sex of an individual
We examined the enamel proteome of two skeletons exhumed from a peculiar archaeological context, exploiting the sex-specificity of AMELY
Summary
A paper by Stewart et al.[7] has revolutionized the way to achieve sex determination of skeletons from archaeological and forensic contexts, thanks to the enamel proteins. One of these proteins is amelogenin, whose gene is translated into two isoforms linked to sexual chromosomes[8], namely AMELX (present in both sexes) and AMELY (restricted to the male sex only), which constitute 90% of the entire enamel proteome[9]. We determined the sex of the so-called ‘Lovers of Modena’, two adult individuals intentionally buried hand-in-hand (Fig. 1; see Figs S17 and[12]) They were found in an Italian Late Antiquity cemetery (4th‒6th century, Ciro Menotti, Modena)[12], together with other eleven individuals. To strengthen the method for sex estimation by ion chromatograms[7], we propose the use of three-peptide peaks, with the aim of confidently identifying AMELY
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