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Structural analysis of Cu/Zn-superoxide dismutase linked to neurodegenerative disease by antibody-guided cryo-EM
Yuki Shino1, Norifumi Muraki1,2, Yui Kobatake3,4
1Department of Chemistry, Keio University, Yokohama, Japan.
Abstract:
Accumulation of misfolded proteins is a hallmark of many neurodegenerative diseases. To characterize such misfolded species in vivo, conformation-specific antibodies are widely used; however, limited knowledge of antibody-epitope interactions often hampers mechanistic insight. To address this, we determined the cryo-electron microscopy structure of the complex between a monoclonal antibody, 19A9, and Cu/Zn-superoxide dismutase (SOD1), a protein associated with canine degenerative myelopathy (DM), which is related to human amyotrophic lateral sclerosis. Biochemical analyses confirmed that 19A9 specifically recognizes monomeric SOD1, and the structure revealed binding near the interface normally used for homodimerization in native SOD1, with steric hindrance preventing interaction when the protein is in its homodimeric form. Immunofluorescence staining of spinal cord sections revealed that 19A9 stained a subset of motoneurons in DM-affected dogs, but not in asymptomatic controls. Structural characterization of the 19A9-monomeric SOD1 complex enabled us to propose that SOD1 monomers can arise in vivo under pathological conditions.
Insights
Researchers characterized the structure of a specific antibody bound to Cu/Zn-superoxide dismutase (SOD1). This revealed how the antibody recognizes misfolded SOD1 monomers, offering insights into neurodegenerative disease mechanisms.
Area of Science:
- Neuroscience
- Structural Biology
- Biochemistry
Background:
- Misfolded proteins accumulate in neurodegenerative diseases.
- Conformation-specific antibodies are used to study these proteins, but their interactions are often poorly understood.
Purpose of the Study:
- To determine the structure of the 19A9 antibody complexed with Cu/Zn-superoxide dismutase (SOD1).
- To understand the antibody-epitope interaction for studying misfolded SOD1 in vivo.
Main Methods:
- Cryo-electron microscopy was used to determine the structure of the 19A9 antibody and SOD1 complex.
- Biochemical analyses confirmed antibody specificity.
- Immunofluorescence staining was performed on spinal cord sections.
Main Results:
- The cryo-EM structure revealed that the 19A9 antibody binds to monomeric SOD1 near its homodimerization interface.
- Binding of the antibody prevents SOD1 homodimerization.
- 19A9 stained a subset of motoneurons in dogs with degenerative myelopathy.
Conclusions:
- The structural and biochemical data elucidate the interaction between 19A9 and monomeric SOD1.
- The findings suggest that SOD1 monomers can form in vivo under pathological conditions, potentially contributing to neurodegeneration.
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