open access publication

Article, 2024

Sequential trypsin and ProAlanase digestions unearth immunological protein biomarkers shrouded by skeletal collagen

iScience, ISSN 2589-0042, Volume 27, 5, Page 109663, 10.1016/j.isci.2024.109663

Contributors

Wilkin, Shevan 0000-0003-0797-6244 (Corresponding author) [1] [2] [3] Lanigan, Liam Thomas 0000-0001-6415-3506 (Corresponding author) [4] Montes, Nuria [5] Sharma, Mukul 0000-0001-8114-0096 [6] Avanzi, Charlotte 0000-0002-1062-4058 [7] Sejdiu, Donikë [3] Majander, Kerttu K 0000-0002-7922-4482 [3] Pfrengle, Saskia [3] [8] Chiang, Yun [4] Kunz, Laura [3] Dittmann, Antje [3] Rühli, Frank Jakobus 0000-0001-7937-001X [3] Singh, Pushpendra 0000-0001-9453-8669 [6] [9] Coll, Maria Fontanals [10] Collins, Matthew James 0000-0003-4226-5501 [4] [11] Taurozzi, Alberto John 0000-0003-0378-1626 (Corresponding author) [4] Schuenemann, Verena J 0000-0002-8593-3672 (Corresponding author) [3]

Affiliations

  1. [1] Griffith University
    2. [NORA names: Australia; Oceania; OECD];
  2. [2] Max Planck Institute of Geoanthropology
    3. [NORA names: Germany; Europe, EU; OECD];
  3. [3] University of Zurich
    4. [NORA names: Switzerland; Europe, Non-EU; OECD];
  4. [4] University of Copenhagen
    5. [NORA names: KU University of Copenhagen; University; Denmark; Europe, EU; Nordic; OECD];
  5. [5] Autonomous University of Barcelona
    6. [NORA names: Spain; Europe, EU; OECD];

Abstract

This study investigates the efficacy of proteomic analysis of human remains to identify active infections in the past through the detection of pathogens and the host response to infection. We advance leprosy as a case study due to the sequestering of sufferers in leprosaria and the suggestive skeletal lesions that can result from the disease. Here we present a sequential enzyme extraction protocol, using trypsin followed by ProAlanase, to reduce the abundance of collagen peptides and in so doing increase the detection of non-collagenous proteins. Through our study of five individuals from an 11th to 18th century leprosarium, as well as four from a contemporaneous non-leprosy associated cemetery in Barcelona, we show that samples from 2 out of 5 leprosarium individuals extracted with the sequential digestion methodology contain numerous host immune proteins associated with modern leprosy. In contrast, individuals from the non-leprosy associated cemetery and all samples extracted with a trypsin-only protocol did not. Through this study, we advance a palaeoproteomic methodology to gain insights into the health of archaeological individuals and take a step toward a proteomics-based method to study immune responses in past populations.

Keywords

Barcelona, abundance, active infection, analysis of human remains, archaeological individuals, biomarkers, case study, cases, cemetery, collagen, collagen peptides, detection, detection of pathogens, digestion, digestion methodology, disease, efficacy, extraction protocol, health, host, host response to infection, human remains, immune response, individuals, infection, leprosarium, leprosy, lesions, method, methodology, non-collagenous proteins, pathogens, peptide, population, protein, protein biomarkers, proteomics-based methods, protocol, remains, response, response to infection, samples, sequential trypsin, sequestering, skeletal lesions, study, study immune responses, suffering, trypsin

Funders

  • Danish National Research Foundation
  • Carlsberg Foundation

Data Provider: Digital Science

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