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Modification of Superoxide Dismutase 1 (SOD1) Properties by a GFP Tag - Implications for Research into Amyotrophic Lateral Sclerosis (ALS)

Stevens, JC; Chia, R; Hendriks, WT; Bros-Facer, V; van Minnen, J; Martin, JE; Jackson, GS; ... Fisher, EMC; + view all (2010) Modification of Superoxide Dismutase 1 (SOD1) Properties by a GFP Tag - Implications for Research into Amyotrophic Lateral Sclerosis (ALS). PLOS ONE , 5 (3) , Article e9541. 10.1371/journal.pone.0009541. Green open access

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Abstract

Background: Since the discovery that mutations in the enzyme SOD1 are causative in human amyotrophic lateral sclerosis (ALS), many strategies have been employed to elucidate the toxic properties of this ubiquitously expressed mutant protein, including the generation of GFP-SOD1 chimaeric proteins for studies in protein localization by direct visualization using fluorescence microscopy. However, little is known about the biochemical and physical properties of these chimaeric proteins, and whether they behave similarly to their untagged SOD1 counterparts.Methodology/Principal Findings: Here we compare the physicochemical properties of SOD1 and the effects of GFP-tagging on its intracellular behaviour. Immunostaining demonstrated that SOD1 alone and GFP-SOD1 have an indistinguishable intracellular distribution in PC12 cells. Cultured primary motor neurons expressing GFP or GFP-SOD1 showed identical patterns of cytoplasmic expression and of movement within the axon. However, GFP tagging of SOD1 was found to alter some of the intrinsic properties of SOD1, including stability and specific activity. Evaluation of wildtype and mutant SOD1, tagged at either the N- or C-terminus with GFP, in PC12 cells demonstrated that some chimaeric proteins were degraded to the individual proteins, SOD1 and GFP.Conclusions/Significance: Our findings indicate that most, but not all, properties of SOD1 remain the same with a GFP tag.

Type: Article
Title: Modification of Superoxide Dismutase 1 (SOD1) Properties by a GFP Tag - Implications for Research into Amyotrophic Lateral Sclerosis (ALS)
Open access status: An open access version is available from UCL Discovery
DOI: 10.1371/journal.pone.0009541
Publisher version: http://dx.doi.org/10.1371/journal.pone.0009541
Language: English
Additional information: © 2010 Stevens et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. This work was supported by research grants from The Ipsen Fund, the UK Motor Neurone Disease Association, the Research Advisory Board of St Bartholomew's and the Royal London Charitable Foundation (RAB04/F7), The Brain Research Trust, the Biotechnology and Biological Sciences Research Council, Cancer Research UK and EU FP6 NEST grant 12702. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
Keywords: FAMILIAL ALS, MUTANT SOD1, HUMAN-CELLS, MUTATIONS, PROTEIN, ULTRACENTRIFUGATION, DEATH, MODEL, GENE, MICE
UCL classification: UCL
UCL > Provost and Vice Provost Offices
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Brain Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Brain Sciences > UCL Institute of Prion Diseases
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Brain Sciences > UCL Institute of Prion Diseases > MRC Prion Unit at UCL
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Brain Sciences > UCL Queen Square Institute of Neurology
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Brain Sciences > UCL Queen Square Institute of Neurology > Department of Neuromuscular Diseases
URI: https://discovery.ucl.ac.uk/id/eprint/126612
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