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MS785-MS27, a Commercially Available Antibody Cocktail against Misfolded SOD1, Recognizes Various Conformation-disordered SOD1 Species Lacking the Incorporated Zn ion
Tokuda, E.; Sakashita, Y.; Tokoro, N.; Date, A.; Kosuge, Y.; Miyasaka, T. MS785-MS27 Reactive Misfolded/Non-Native Zn-Deficient SOD1 Species Exhibit Cytotoxicity and Adopt Heterozygous Conformations in Motor Neurons. Int. J. Mol. Sci.2024, 25, 5603.
Tokuda, E.; Sakashita, Y.; Tokoro, N.; Date, A.; Kosuge, Y.; Miyasaka, T. MS785-MS27 Reactive Misfolded/Non-Native Zn-Deficient SOD1 Species Exhibit Cytotoxicity and Adopt Heterozygous Conformations in Motor Neurons. Int. J. Mol. Sci. 2024, 25, 5603.
Tokuda, E.; Sakashita, Y.; Tokoro, N.; Date, A.; Kosuge, Y.; Miyasaka, T. MS785-MS27 Reactive Misfolded/Non-Native Zn-Deficient SOD1 Species Exhibit Cytotoxicity and Adopt Heterozygous Conformations in Motor Neurons. Int. J. Mol. Sci.2024, 25, 5603.
Tokuda, E.; Sakashita, Y.; Tokoro, N.; Date, A.; Kosuge, Y.; Miyasaka, T. MS785-MS27 Reactive Misfolded/Non-Native Zn-Deficient SOD1 Species Exhibit Cytotoxicity and Adopt Heterozygous Conformations in Motor Neurons. Int. J. Mol. Sci. 2024, 25, 5603.
Abstract
Misfolding of superoxide dismutase-1 (SOD1) is a pathological hallmark of amyotrophic lateral sclerosis (ALS) with SOD1 mutations. The development of antibodies specific for misfolded SOD1 deepens our understanding of how the protein participates in ALS pathogenesis. Since the term “misfolding” refers to various disordered conformers other than the natively folded one, which misfolded species are recognized by specific antibodies should be determined. Here, we molecularly characterized the recognition by MS785-MS27, an antibody cocktail experimentally confirmed to recognize over 100 ALS-linked SOD1 mutants. Indirect ELISA revealed that the antibody cocktail recognized Zn-deficient wild-type and mutated SOD1 species. It also recognized conformation-disordered wild-type and mutated SOD1 species, such as unfolded and oligomeric forms, but had less affinity for the aggregated form. Antibody-reactive SOD1 exhibited cytotoxicity to a motor neuron cell model, which was blocked by Zn treatment with Zn-deficient SOD1. Immunohistochemistry revealed antibody-reactive SOD1 mainly in spinal motor neurons of SOD1G93A mice throughout the disease course, and the distribution after symptomatic stages differed from that of other misfolded SOD1 species. This suggested that misfolded SOD1 species exist as heterogeneous populations. In conclusion, MS785-MS27 recognizes various misfolded SOD1 lacking the incorporated Zn ion and should be useful for monitoring Zn availability in SOD1.
Biology and Life Sciences, Biochemistry and Molecular Biology
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