2-DE and MALDI-TOF-MS for a comparative analysis of proteins expressed in different cellular models of amyotrophic lateral sclerosis.
Amyotrophic lateral sclerosis (ALS) is a fatal, neurodegenerative disorder characterized by the selective loss of motor neurons from the spinal cord and brain. About 10% of ALS cases are familial (FALS), and in 20% of these cases the disease has been linked to mutations in the Cu,Zn-SOD1 gene. Although the molecular mechanisms causing these forms of ALS are still unclear, evidence has been provided that motor neurons injuries associated with mutant superoxide dismutase (SOD1)-related FALS result from a toxic gain-in-fuction of the mutated enzyme. To understand better the role of these mutations in the pathophysiology of FALS we have compared the pattern of proteins expressed in human neuroblastoma SH-SY5Y cell line with those of cell lines transfected with plasmids expressing the wild-type human SOD1 and the H46R and G93A mutants. 2-DE coupled to MALDI-TOF-MS were the proteomic tools used for identification of differentially expressed proteins. These included cytoskeletal proteins, proteins that regulate energetic metabolism and intracellular redox conditions, and the ubiquitin proteasome system. The proteomic approach allowed to expand the knowledge on the pattern of proteins, with altered expression, which we should focus on, for a better understanding of the possible mechanism involved in mutated-SOD1 toxicity. The cellular models considered in this work have also evidenced biochemical characteristics common to other SOD1-mutated cellular lines connected to the pathogenesis of ALS.
Di Poto
Cristina
C
Department of Biochemistry A Castellani, University of Pavia, Pavia, Italy.
Iadarola
Paolo
P
Bardoni
Anna Maria
AM
Passadore
Ileana
I
Giorgetti
Sofia
S
Cereda
Cristina
C
Carrì
Maria Teresa
MT
Ceroni
Mauro
M
Salvini
Roberta
R
D019943:Amino Acid Substitution; D000690:Amyotrophic Lateral Sclerosis / Q000378:metabolism*; D017209:Apoptosis; D002457:Cell Extracts / Q000032:analysis; D002460:Cell Line; D002478:Cells, Cultured; D003600:Cytosol / Q000737:chemistry*; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D004789:Enzyme Activation / Q000235:genetics; D006801:Humans; D008954:Models, Biological; D009447:Neuroblastoma / Q000378:metabolism; D016900:Neurofilament Proteins / Q000172:deficiency / Q000378:metabolism; D010084:Oxidation-Reduction; D011506:Proteins / Q000032:analysis* / Q000145:classification; D020543:Proteome / Q000032:analysis; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D013116:Spinal Cord / Q000166:cytology / Q000473:pathology; D013482:Superoxide Dismutase / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D014162:Transfection
D002457:Cell Extracts; D016900:Neurofilament Proteins; D011506:Proteins; D020543:Proteome; C109415:SOD1 G93A protein; D013482:Superoxide Dismutase