Sequential trypsin and ProAlanase digestions unearth immunological protein biomarkers shrouded by skeletal collagen
Shevan Wilkin,
Liam T. Lanigan,
Nuria Montes,
Mukul Sharma,
Charlotte Avanzi,
Donikë Sejdiu,
Kerttu Majander,
Saskia Pfrengle,
Yun Chiang,
Laura Kunz,
Antje Dittmann,
Frank Rühli,
Pushpendra Singh,
Maria Fontanals Coll,
Matthew J. Collins,
Alberto J. Taurozzi,
Verena J. Schuenemann
Affiliations
Shevan Wilkin
Institute of Evolutionary Medicine, University of Zurich, Zurich, Switzerland; Max Planck Institute of Geoanthropology, Jena, Germany; Australian Research Centre for Human Evolution, Griffith University, Brisbane, QLD, Australia; Corresponding author
Liam T. Lanigan
Globe Institute, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark; Corresponding author
Nuria Montes
Unitat d'Antropologia Biològica, Departament de Biologia Animal, Biologia Vegetal i Ecologia, Universitat Autònoma de Barcelona, Barcelona, Spain
Mukul Sharma
Microbial Pathogenesis and Genomics, National Institute of Research in Tribal Health, Jabalpur, MP, India
Charlotte Avanzi
Department of Microbiology, Immunology, and Pathology, Colorado State University, Fort Collins, CO, USA
Donikë Sejdiu
Institute of Evolutionary Medicine, University of Zurich, Zurich, Switzerland
Kerttu Majander
Institute of Evolutionary Medicine, University of Zurich, Zurich, Switzerland
Saskia Pfrengle
Institute of Evolutionary Medicine, University of Zurich, Zurich, Switzerland; Institute for Archaeological Sciences, University of Tübingen, Tübingen, Germany
Yun Chiang
Globe Institute, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark
Laura Kunz
Functional Genomics Center Zurich, University of Zurich, Zurich, Switzerland
Antje Dittmann
Functional Genomics Center Zurich, University of Zurich, Zurich, Switzerland
Frank Rühli
Institute of Evolutionary Medicine, University of Zurich, Zurich, Switzerland
Pushpendra Singh
Microbial Pathogenesis and Genomics, National Institute of Research in Tribal Health, Jabalpur, MP, India; Model Rural Health Research Unit, Badoni, Datia (MP), India
Maria Fontanals Coll
BioArCh - Department of Archaeology, University of York, York, UK
Matthew J. Collins
Globe Institute, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark; McDonald Institute for Archaeological Research, University of Cambridge, West Tower, Downing St, Cambridge CB2 3ER, UK
Alberto J. Taurozzi
Globe Institute, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark; Corresponding author
Verena J. Schuenemann
Institute of Evolutionary Medicine, University of Zurich, Zurich, Switzerland; Corresponding author
Summary: 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.
