Dissection of schistosome tissues under LC-MS compatible preservative conditions for quantitative proteomics

Neves, Leandro Xavier ORCID: 0000-0002-6074-1025, Wilson, R Alan, Brownridge, Philip, Holman, Stephen WW, Harman, Victoria MM ORCID: 0000-0002-0990-153X, Eyers, Claire EE ORCID: 0000-0002-3223-5926, Beynon, Robert JJ ORCID: 0000-0003-0857-495X and Castro-Borges, William (2023) Dissection of schistosome tissues under LC-MS compatible preservative conditions for quantitative proteomics. RAPID COMMUNICATIONS IN MASS SPECTROMETRY, 38 Sup (S1). e9523-.

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Abstract

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.

Item Type:	Article
Uncontrolled Keywords:	Animals, Schistosoma mansoni, Proteome, Chromatography, Liquid, Proteomics, Tandem Mass Spectrometry, Liquid Chromatography-Mass Spectrometry
Divisions:	Faculty of Health and Life Sciences Faculty of Health and Life Sciences > Institute of Systems, Molecular and Integrative Biology
Depositing User:	Symplectic Admin
Date Deposited:	20 Jun 2023 08:43
Last Modified:	05 Feb 2024 03:17
DOI:	10.1002/rcm.9523
Open Access URL:	https://doi.org/10.1002/rcm.9523
Related URLs:	Author Publisher
URI:	https://livrepository.liverpool.ac.uk/id/eprint/3171121