title: Different immunoreactivity against monoclonal antibodies between wild-type and mutant copper/zinc superoxide dismutase linked to amyotrophic lateral sclerosis.
abstract: Although more than 100 mutations have been identified in the copper/zinc superoxide dismutase (Cu/Zn-SOD) in familial amyotrophic lateral sclerosis (FALS), the mechanism responsible for FALS remains unclear. The finding of the present study shows that FALS-causing mutant Cu/Zn-SOD proteins (FALS mutant SODs), but not wild-type SOD, are barely detected by three monoclonal antibodies (mAbs) in Western blot analyses. The enzyme-linked immunosorbent assay for denatured FALS mutant SODs by dithiothreitol, SDS, or heat treatment also showed a lowered immunoreactivity against the mAbs compared with wild-type SOD. Because all the epitopes of these mAbs are mapped within the Greek key loop (residues 102-115 in human Cu/Zn-SOD), these data suggest that different conformational changes occur in the loop between wild-type and FALS mutant SODs during the unfolding process. Circular dichroism measurements revealed that the FALS mutant SODs are sensitive to denaturation by dithiothreitol, SDS, or heat treatment, but these results do not completely explain the different recognition by the mAbs between wild-type and FALS mutant SODs under the denatured conditions. The study on the conformational changes in local areas monitoring with mAbs may provide a new insight into the etiology of FALS.
authors.affiliation: Department of Biochemistry, Hyogo College of Medicine, 1-1 Mukogawa-cho, Nishinomiya, Hyogo 663-8501, Japan.
authors.forename: Noriko
authors.initials: N
authors.lastname: Fujiwara
authors.forename: Yasuhide
authors.initials: Y
authors.lastname: Miyamoto
authors.forename: Kyoko
authors.initials: K
authors.lastname: Ogasahara
authors.forename: Motoko
authors.initials: M
authors.lastname: Takahashi
authors.forename: Takahisa
authors.initials: T
authors.lastname: Ikegami
authors.forename: Rina
authors.initials: R
authors.lastname: Takamiya
authors.forename: Keiichiro
authors.initials: K
authors.lastname: Suzuki
authors.forename: Naoyuki
authors.initials: N
authors.lastname: Taniguchi
mesh_terms: D000595:Amino Acid Sequence; D000690:Amyotrophic Lateral Sclerosis / Q000276:immunology / Q000378:metabolism*; D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry*; D015153:Blotting, Western; D002460:Cell Line; D002942:Circular Dichroism; D004229:Dithiothreitol / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D051379:Mice; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016296:Mutagenesis; D009154:Mutation; D010455:Peptides / Q000737:chemistry; D012967:Sodium Dodecyl Sulfate / Q000737:chemistry; D013482:Superoxide Dismutase / Q000737:chemistry*; D014466:Ultraviolet Rays
chemical_list: D000911:Antibodies, Monoclonal; D010455:Peptides; D012967:Sodium Dodecyl Sulfate; D013482:Superoxide Dismutase; D004229:Dithiothreitol