title: In-cell NMR reveals potential precursor of toxic species from SOD1 fALS mutants.
abstract: Mutations in the superoxide dismutase 1 (SOD1) gene are related to familial cases of amyotrophic lateral sclerosis (fALS). Here we exploit in-cell NMR to characterize the protein folding and maturation of a series of fALS-linked SOD1 mutants in human cells and to obtain insight into their behaviour in the cellular context, at the molecular level. The effect of various mutations on SOD1 maturation are investigated by changing the availability of metal ions in the cells, and by coexpressing the copper chaperone for SOD1, hCCS. We observe for most of the mutants the occurrence of an unstructured SOD1 species, unable to bind zinc. This species may be a common precursor of potentially toxic oligomeric species, that are associated with fALS. Coexpression of hCCS in the presence of copper restores the correct maturation of the SOD1 mutants and prevents the formation of the unstructured species, confirming that hCCS also acts as a molecular chaperone.
authors.affiliation: 1] CERM, Magnetic Resonance Center, University of Florence, Via Luigi Sacconi 6, Sesto Fiorentino, 50019 Florence, Italy [2] Department of Biomedical, Clinical and Experimental Sciences, University of Florence, Viale Morgagni 50, 50134 Florence, Italy.
authors.forename: Enrico
authors.initials: E
authors.lastname: Luchinat
authors.affiliation: CERM, Magnetic Resonance Center, University of Florence, Via Luigi Sacconi 6, Sesto Fiorentino, 50019 Florence, Italy.
authors.forename: Letizia
authors.initials: L
authors.lastname: Barbieri
authors.affiliation: CERM, Magnetic Resonance Center, University of Florence, Via Luigi Sacconi 6, Sesto Fiorentino, 50019 Florence, Italy.
authors.forename: Jeffrey T
authors.initials: JT
authors.lastname: Rubino
authors.affiliation: Giotto Biotech S.r.l., Via Madonna del Piano 6, Sesto Fiorentino, 50019 Florence, Italy.
authors.forename: Tatiana
authors.initials: T
authors.lastname: Kozyreva
authors.affiliation: 1] CERM, Magnetic Resonance Center, University of Florence, Via Luigi Sacconi 6, Sesto Fiorentino, 50019 Florence, Italy [2] Department of Chemistry, University of Florence, Via della Lastruccia 3, Sesto Fiorentino, 50019 Florence, Italy.
authors.forename: Francesca
authors.initials: F
authors.lastname: Cantini
authors.affiliation: 1] CERM, Magnetic Resonance Center, University of Florence, Via Luigi Sacconi 6, Sesto Fiorentino, 50019 Florence, Italy [2] Department of Chemistry, University of Florence, Via della Lastruccia 3, Sesto Fiorentino, 50019 Florence, Italy [3] Fondazione Farmacogenomica FiorGen onlus, Via L. Sacconi 6, Sesto Fiorentino, 50019 Florence, Italy.
authors.forename: Lucia
authors.initials: L
authors.lastname: Banci
mesh_terms: D000690:Amyotrophic Lateral Sclerosis / Q000235:genetics* / Q000378:metabolism; D003300:Copper / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D018832:Molecular Chaperones / Q000378:metabolism*; D019906:Nuclear Magnetic Resonance, Biomolecular; D017510:Protein Folding; D013482:Superoxide Dismutase / Q000235:genetics* / Q000378:metabolism; D000072105:Superoxide Dismutase-1; D015032:Zinc / Q000378:metabolism
chemical_list: C114678:CCS protein, human; D018832:Molecular Chaperones; C000606290:SOD1 protein, human; D003300:Copper; D013482:Superoxide Dismutase; D000072105:Superoxide Dismutase-1; D015032:Zinc