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Increasing sustainability in palaeoproteomics by optimizing digestion times for large-scale archaeological bone analyses

ABSTRACT: # Description of data generation and experimental procedure Six bones from La Draga (Spain, Holocene, samples LD_01 to LD_06) and Baiyisha Karst Cave (China, Pleistocene, samples BKC_07 to BKC_12) were sampled for the purpose of this study. Initial sampling was divided into three sub-samples for the three digestion duration tested here (site code_sample number_3h, site code_sample number_6h and site code_sample number_18h). Samples were then processed according to the ZooMS protocol: they were demineralised in 0.6 M hydrochloric acid (HCl) for 24 hours. The HCl supernatant was then removed and samples were rinsed thrice in 100 μL ammonium bicarbonate (50 mM, NH4HCO3, hereafter AmBic) for subsequent gelatinisation in a final volume of 100 μL AmBic for one hour at 65C. Following gelatinisation, the 100 μL AmBic solution was transferred to a new microtube, to which 0.8 μg trypsin (Promega) was added for incubation at 37 °C, with mild agitation at 300 rpm (VWR, Thermal Shake lite). Digestion occurred for either 3, 6, or 18 hours. To stop trypsin digestion, 2 μL of 5% trifluoroacetic acid (TFA) was added to each sample. The digested extracts were then split in two parts for separate analyses via matrix-assisted laser desorption/ionisation-time of flight mass spectrometry (MALDI-ToF MS) and liquid-chromatography tandem mass spectrometry (LC-MS/MS). To assess any potential contamination by non-endogenous peptides, we performed extraction of laboratory blanks alongside the samples for each enzymatic digestion condition.

INSTRUMENT(S): Orbitrap Exploris 480

ORGANISM(S): Bovinae

TISSUE(S): Bone

SUBMITTER: Frido Welker

LAB HEAD: Frido Welker

PROVIDER: PXD045027 | Pride | 2024-03-14

REPOSITORIES: Pride

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Increasing sustainability in palaeoproteomics by optimizing digestion times for large-scale archaeological bone analyses

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