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Evidence of glutathione transferase complexing and signaling in the model nematode Caenorhabditis elegans using a pull‐down proteomic assay
Author(s) -
Greetham Darren,
Morgan Charly,
Campbell Alison M.,
van Rossum Arjan J.,
Barrett John,
Brophy Peter M.
Publication year - 2004
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.200300719
Subject(s) - glutathione s transferase , glutathione , proteomics , glutathione transferase , biology , signal transduction , microbiology and biotechnology , chemistry , biochemistry , computational biology , enzyme , gene
Phage display techniques using random peptide interactions have supported the role of mammalian glutathione transferase (GST) as part of a signalling pathway for both oxidative stress and an apoptosis pathway. Little is known about the interaction of nonmammalian GST with other proteins. GSTs have been implicated in the development of chronic nematode infections by neutralising cytotoxic products arising from host immune initiated reactive oxygen species (ROS) assault. In this study we attached one of the key GSTs expressed in the model nematode Caenorhabditis elegans to an affinity support matrix and directly identified major interacting proteins by two‐dimensional electrophoresis and peptide mass fingerprinting before and following oxidative stress. Nematode GST does not appear to be a stand‐alone enzyme and interacts with many types of proteins in both normal and ROS stress conditions. Pull‐down proteomic presents a flexible, label free, rapid and economical assay without specialised ligand fishing equipment to identify protein binding partners.