Dissection of schistosome tissues under LC–MS compatible preservative conditions for quantitative proteomics.
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2023
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Schistosomes are blood flukes with specialised tissues and organs, each one playing a
pivotal role in perpetuating the parasite life cycle. Herein, we describe a detailed
methodology for preserving the proteome of adult Schistosoma mansoni worms during
manual dissection for enrichment of tissues associated with the parasite's alimentary
tract. We provide step-by-step directions for specimen storage and dissection while in
preservative solution, tissue homogenisation, protein extraction and digestion using a
methodology fully compatible with downstream quantitative liquid chromatography–
mass spectrometry analysis. Our methodology uses label-free and QconCAT-based
absolute quantification for detection of S. mansoni oesophageal gland products
proposed as vaccine candidates. Through stabilisation of the proteome and minimising
sample degradation during dissection our approach has allowed us to access the
hidden proteome of target tissues not readily available from total lysates because of
their small volume. This protocol can be replicated or adapted to other Schistosoma
species lacking quantitative proteomics characterisation of specialised tissues for
discovery of proteins with potential diagnostic and therapeutic utility.
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NEVES, L. X. et al. Dissection of schistosome tissues under LC–MS compatible preservative conditions for quantitative proteomics. Rapid Communications in Mass Spectrometry, artigo e9523, 2023. Disponível em: <https://analyticalsciencejournals.onlinelibrary.wiley.com/doi/10.1002/rcm.9523>. Acesso em: 01 ago. 2023.