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Paleoproteomic study of the Iceman’s brain tissue

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The Tyrolean Iceman, a Copper-age ice mummy, is one of the best-studied human individuals. While the genome of the Iceman has largely been decoded, tissue-specific proteomes have not yet been investigated. We studied the proteome of two distinct brain samples using gel-based and liquid chromatography–mass spectrometry-based proteomics technologies together with a multiple-databases and -search algorithms-driven data-analysis approach. Thereby, we identified a total of 502 different proteins. Of these, 41 proteins are known to be highly abundant in brain tissue and 9 are even specifically expressed in the brain. Furthermore, we found 10 proteins related to blood and coagulation. An enrichment analysis revealed a significant accumulation of proteins related to stress response and wound healing. Together with atomic force microscope scans, indicating clustered blood cells, our data reopens former discussions about a possible injury of the Iceman’s head near the site where the tissue samples have been extracted.

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The work was supported in part by the SüdtirolerSparkasse. F.M., G.G. and A.Z. were supported by the law 14 grant of the province Bolzano, South Tyrol, Italy. A.T., T.O. and D.L. were supported by the Cluster of Excellence “Inflammation at Interfaces”. B.V.D.B. was supported by the SFB877-project Z2.

Conflict of interest

A.K. and M.J. are affiliates of Siemens Healthcare, Erlangen, Germany.

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Correspondence to Andreas Keller.

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F. Maixner, T. Overath, and D. Linke contributed equally as first authors.

A. Tholey, A. Zink and A. Keller contributed equally as senior authors.

Electronic supplementary material

The MS -raw data as well as database search results (Filename: “Protein_Results.xlsx”) are accessible at: “” under the login: “sukmb289b” and the password: “eiwai9ce”. Below is the link to the electronic supplementary material.

Supplementary material 4 (DOCX 47078 kb)


Supplementary material 1 An overview of the proteins identified in-solution and in-gel for both samples, 1024 and 1025, respectively. (XLSX 71 kb)


Supplementary material 2 The significant pathways for functional modelling (KEGG and gene ontologies) after adjusting with the Benjamini–Hochberg approach. (XLSX 63 kb)


Supplementary material 3 A multi-tab table file with information regarding protein identification by the four search engines applied (Mascot, SEQUEST, OMSSA and X!Tandem) either (1) with specification of a protease (Trypsin) or (2) without specification of a protease (“No Enzyme”), respectively. For the in-solution analyses, the identified proteins are listed regarding the number of matches by the four search engines. For the samples analyzed after gel separation, the information about protein identification is linked to the position in the gel, (1) for each of the four search engines (non-merged data) and (2) after merging the data into a form representing a picture of the gel. (XLSX 1966 kb)

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Maixner, F., Overath, T., Linke, D. et al. Paleoproteomic study of the Iceman’s brain tissue. Cell. Mol. Life Sci. 70, 3709–3722 (2013).

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