ИСТИНА

Determining the taxonomic affiliation and sex of animals based on fossil bone remains is one of the most important questions in studying the lifestyle of ancient people. The use of traditional morphological methods based on the analysis of skeletal markers of sex differences is often difficult due to the high degree of fragmentation of bone remains and the lack of morphological markers. Determining the species belonging by fragmentary bone remains in addition to traditional morphological methods, are currently effectively solved by the methods of genetic analysis and Zooarchaeology by Mass Spectrometry analysis (ZooMS). Another method for determining sex is the proteomic analysis of sex-specific amelogenin peptides in tooth enamel. Protein is considerably more robust than DNA molecules, particularly when it is physically associated with mineral surfaces such as tooth enamel. Therefore, it is the most appropriate approach for biochemical sex determination. This method was developed and successfully applied in the study of sex in archaeological skeletons of people of different antiquity and taphonomic preservation, both adults and children. The investigation was conducted on the series of teeth of modern bisons with known sexs and age, and several groups of ancient bisons dated to Paleolithic period. Etching of tooth enamel and extraction of peptides were performed according to the modified technique developed in previous studies of human teeth. Liquid chromatography and mass spectrometry was performed with an Ultimate 3000 Nano LC System which was coupled to the Orbitrap Q Exactive Plus mass spectrometer via a nanoelectrospray source. MS/MS data were analyzed with the Peaks studio 10 (Bioinformatics Solutions Inc.). Peptide sequences generated by the PEAKS Studio were analyzed using the UniProtKB/Swiss-Prot (25.10.2019) database for Bos taurus (Bovine) as a reference. The list of identified proteins contains proteins for which at least two unique peptide sequences were determined. On a control group of bison with known sex and age, the analysis conducted by blind test showed its effectiveness. The effectiveness on Bovinae is identical to the effectiveness of the same method for human samples. The presence of AMEX and AMELY genes in other animal species suggests the possibility of conducting a peptidomic analysis for sex determine. The method has also demonstrated its effectiveness on archaeological materials dated to Late Pleistocene. Sex determination is possible and for the permanent and deciduous teeth. Taking into account the fact that bison were often the main life-supporting strategies of the oldest Pleistocene populations, it is difficult to overestimate the prospects of this method for zooarchaeological and archaeological research.