Optimizing separation efficiency of 2-DE procedures for visualization of different superoxide dismutase forms in a cellular model of amyotrophic lateral sclerosis.
Neurodegenerative diseases such as Alzheimer disease (AD) and Parkinson disease (PD) have been associated with increased production of reactive oxygen species. In AD and PD patients, superoxide dismutase (SOD1) was also indicated as a major target of oxidative damage. In particular, in brain tissue of these patients, different SOD1 isoforms have been identified, although their functional role still remains to be elucidated. In the light of the possibility that different SOD1 entities could be expressed also in other neurodegenerative disorders, as a sort of unifying event with AD and PD, we have investigated amyotrophic lateral sclerosis (ALS) using human neuroblastoma SH-SY5Y cells with mutated SOD1 gene H46R as cellular model. 2-DE using a narrow-range IPG 4-7 strips in the first dimension and linear 15% SDS-PAGE in the second allowed to separate different SOD1 spots. MALDI-TOF MS and CapLC-MS/MS have been used for their complete identification. This is the first report in which the presence of SOD1 (iso) forms in a cellular model of ALS has been evidenced.
Di Poto
Cristina
C
Department of Biochemistry, University of Pavia, Pavia, Italy.
Iadarola
Paolo
P
Salvini
Roberta
R
Passadore
Ileana
I
Cereda
Cristina
C
Ceroni
Mauro
M
Bardoni
Anna Maria
AM
D000690:Amyotrophic Lateral Sclerosis / Q000201:enzymology*; D053801:Capillary Electrochromatography / Q000379:methods; D002457:Cell Extracts / Q000032:analysis / Q000737:chemistry; D003600:Cytosol / Q000201:enzymology*; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D006801:Humans; D007527:Isoenzymes / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D008956:Models, Chemical; D009154:Mutation; D009447:Neuroblastoma / Q000201:enzymology; D010084:Oxidation-Reduction; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D013482:Superoxide Dismutase / Q000737:chemistry* / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D014162:Transfection; D014407:Tumor Cells, Cultured
D002457:Cell Extracts; D007527:Isoenzymes; D013482:Superoxide Dismutase