University of Tasmania
Browse

File(s) under permanent embargo

Sequential protein extraction as an efficient method for improved proteome coverage in larvae of Atlantic salmon (Salmo salar)

journal contribution
posted on 2023-05-18, 20:18 authored by Nuez-Ortin, W, Christopher CarterChristopher Carter, Nichols, PD, Richard WilsonRichard Wilson
Understanding diet- and environmentally induced physiological changes in fish larvae is a major goal for the aquaculture industry. Proteomic analysis of whole fish larvae comprising multiple tissues offers considerable potential but is challenging due to the very large dynamic range of protein abundance. To extend the coverage of the larval phase of the Atlantic salmon (Salmo salar) proteome, we applied a two-step sequential extraction (SE) method, based on differential protein solubility, using a nondenaturing buffer containing 150 mM NaCl followed by a denaturing buffer containing 7 M urea and 2 M thiourea. Extracts prepared using SE and one-step direct extraction were characterized via label-free shotgun proteomics using nanoLC-MS/MS (LTQ-Orbitrap). SE partitioned the proteins into two fractions of approximately equal amounts, but with very distinct protein composition, leading to identification of ∼40% more proteins than direct extraction. This fractionation strategy enabled the most detailed characterization of the salmon larval proteome to date and provides a platform for greater understanding of physiological changes in whole fish larvae. The MS data are available via the ProteomeXchange Consortium PRIDE partner repository, dataset PXD003366.

History

Publication title

Proteomics

Volume

16

Issue

14

Pagination

2043-2047

ISSN

1615-9861

Department/School

Institute for Marine and Antarctic Studies

Publisher

WILEY-VCH Verlag GmbH & Co. KGaA

Place of publication

Weinheim, Germany

Rights statement

Copyright 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim

Repository Status

  • Restricted

Socio-economic Objectives

Fisheries - aquaculture not elsewhere classified

Usage metrics

    University Of Tasmania

    Exports

    RefWorks
    BibTeX
    Ref. manager
    Endnote
    DataCite
    NLM
    DC