High-Resolution NMR H/D Exchange of Human Superoxide Dismutase Inclusion Bodies Reveals Significant Native Features Despite Structural Heterogeneity.
Dalia NaserMichael V TarascaBruna SiebeneichlerAnna SchaeferHarmeen K DeolTyler G B SouleJohnathan AlmeySusan KelsoGyana G MishraHilary SimonElizabeth M MeieringPublished in: Angewandte Chemie (International ed. in English) (2022)
Protein aggregation is central to aging, disease and biotechnology. While there has been recent progress in defining structural features of cellular protein aggregates, many aspects remain unclear due to heterogeneity of aggregates presenting obstacles to characterization. Here we report high-resolution analysis of cellular inclusion bodies (IBs) of immature human superoxide dismutase (SOD1) mutants using NMR quenched amide hydrogen/deuterium exchange (qHDX), FTIR and Congo red binding. The extent of aggregation is correlated with mutant global stability and, notably, the free energy of native dimer dissociation, indicating contributions of native-like monomer associations to IB formation. This is further manifested by a common pattern of extensive protection against H/D exchange throughout nine mutant SOD1s despite their diverse characteristics. These results reveal multiple aggregation-prone regions in SOD1 and illuminate how aggregation may occur via an ensemble of pathways.
Keyphrases
- high resolution
- endothelial cells
- single cell
- amyotrophic lateral sclerosis
- mass spectrometry
- induced pluripotent stem cells
- binding protein
- wild type
- pluripotent stem cells
- protein protein
- amino acid
- tandem mass spectrometry
- genome wide
- high speed
- small molecule
- dna methylation
- convolutional neural network
- nitric oxide
- visible light