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In vitro evidence consistent with an interaction between wild‐type and mutant SOD 1 protein associated with canine degenerative myelopathy
Author(s) -
Qi Yao,
Montague Paul,
Loney Colin,
Campbell Clare,
Shafie Intan N. F.,
Anderson Thomas J.,
McLaughlin Mark
Publication year - 2019
Publication title -
european journal of neuroscience
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.346
H-Index - 206
eISSN - 1460-9568
pISSN - 0953-816X
DOI - 10.1111/ejn.14526
Subject(s) - mutant , sod1 , superoxide dismutase , wild type , biology , fusion protein , in vitro , amyotrophic lateral sclerosis , transfection , mutant protein , green fluorescent protein , microbiology and biotechnology , mutation , gene , genetics , biochemistry , enzyme , pathology , medicine , recombinant dna , disease
Canine degenerative myelopathy ( DM ) is a progressive neurological disorder that may be considered to be a large animal model for specific forms of the fatal human disease, familial amyotrophic lateral sclerosis ( fALS ). DM is associated with a c118G>A mutation of the superoxide dismutase 1 ( Sod 1) gene, and a significant proportion of cases are inherited in an autosomal recessive manner in contrast to the largely, but not exclusively, dominant mode of inheritance in fALS . The consensus view is that these Sod 1/ SOD 1 mutations result in a toxic gain of function but the mechanisms remain unclear. Here we used an in vitro neuroblastoma cell line transfection system to monitor wild‐type and mutant forms of SOD 1 fusion proteins containing either a Cherry or an enhanced green fluorescent protein ( EGFP ) tag. These fusion proteins retained SOD 1 enzymatic activity on a native gel assay system. We demonstrate that SOD 1 aggregate density is significantly higher in DM transfectants compared to wild‐type. In addition, we show by co‐immunoprecipitation and confocal microscopy, evidence for a potential interaction between wild‐type and mutant forms of SOD1 in co‐transfected cells. While in vitro studies have shown SOD 1 heterodimer formation in fALS models, this is the first report for DM SOD 1. Therefore, despite for the majority of cases there is a difference in the mode of inheritance between fALS and DM , a similar interaction between wild‐type and mutant SOD 1 forms can occur. Clarifying the role of SOD 1 in DM may also be of benefit to understanding the role of SOD 1 in fALS .