Evidence of glutathione transferase complexing and signaling in the model nematode Caenorhabditis elegans using a pull-down proteomic assay.
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.
Greetham
Darren
D
Institute of Biological Sciences, University of Wales, Aberystwyth SY23 3DA, Wales, UK.
Morgan
Charly
C
Campbell
Alison M
AM
van Rossum
Arjan J
AJ
Barrett
John
J
Brophy
Peter M
PM
D000595:Amino Acid Sequence; D000818:Animals; D017173:Caenorhabditis elegans; D019992:Databases as Topic; D015180:Electrophoresis, Gel, Two-Dimensional; D005982:Glutathione Transferase / Q000737:chemistry / Q000378:metabolism*; D006861:Hydrogen Peroxide / Q000378:metabolism; D013058:Mass Spectrometry; D008954:Models, Biological; D008969:Molecular Sequence Data; D040901:Proteomics / Q000379:methods*; D017382:Reactive Oxygen Species; D011994:Recombinant Proteins / Q000737:chemistry; D015398:Signal Transduction; D012965:Sodium Chloride / Q000494:pharmacology; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014357:Trypsin / Q000494:pharmacology
D017382:Reactive Oxygen Species; D011994:Recombinant Proteins; D012965:Sodium Chloride; D006861:Hydrogen Peroxide; D005982:Glutathione Transferase; D014357:Trypsin