Journal of Proteomics ( IF 2.8 ) Pub Date : 2020-11-02 , DOI: 10.1016/j.jprot.2020.104039 Georgia Ntasi 1 , Daniel P Kirby 2 , Ilaria Stanzione 1 , Andrea Carpentieri 3 , Patrizia Somma 4 , Paola Cicatiello 1 , Gennaro Marino 3 , Paola Giardina 1 , Leila Birolo 1
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Identification and characterization of ancient proteins still require technical developments towards non-invasiveness, sensitivity, versatility and ease of use of the analyses. We report that the enzyme functionalized films, described in Cicatiello et al. (2018), can be used efficiently on the surface of different objects ranging from fixative-coated paper to canvas to the coating on an albumen photograph, as well as the much harder surfaces of ivory objects and the proteinaceous binders in the decoration of a wooden Egyptian coffin. The mixture of digested peptides that are efficiently captured on the functionalized surface are also amenable to LC-MS/MS analysis, which is necessary to confidently identify chemical modifications induced upon degradation, in order to characterize the conservation state of proteins. Moreover, in a two-step procedure, we have combined the trypsin functionalized film with a PNGaseF functionalized film, which adds a deglycosylation pretreatment allowing improved detection of glycosylated proteins.
Significance
User friendly trypsin functionalized films were implemented to expand their potential as versatile, modular tools that can be widely exploited in the world of diagnosis of cultural heritage objects, ancient proteins, and palaeoproteomics: a procedure that could be carried out by conservators or archaeologists first on-site and later analysed with standard MS techniques.
中文翻译：
一种通用且用户友好的方法，用于分析古代和历史物体中的蛋白质
古代蛋白质的鉴定和表征仍然需要向分析的非侵入性、敏感性、多功能性和易用性发展。我们报告了 Cicatiello 等人描述的酶功能化薄膜。(2018)，可以有效地用于不同物体的表面，从固定剂涂层纸到帆布再到蛋白照片上的涂层，以及象牙物体的坚硬表面和木质装饰中的蛋白质粘合剂埃及棺材。在功能化表面上有效捕获的消化肽混合物也适用于 LC-MS/MS 分析，这是自信地识别降解后诱导的化学修饰所必需的，以表征蛋白质的保守状态。此外，在两步过程中，
意义
实施了用户友好的胰蛋白酶功能化薄膜，以扩展其作为多功能模块化工具的潜力，可在文化遗产对象、古代蛋白质和古蛋白质组学的诊断领域广泛利用：保护人员或考古学家首先可以执行的程序位点，然后用标准 MS 技术进行分析。
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