title: Local unfolding in a destabilized, pathogenic variant of superoxide dismutase 1 observed with H/D exchange and mass spectrometry.
abstract: Hydrogen exchange monitored by mass spectrometry has been used to study the structural behavior of the pathogenic A4V variant of superoxide dismutase 1 (SOD1) in the metal-free (apo) form. Mass spectrometric data revealed that in the disulfide-intact (S-S) form, the A4V variant is destabilized at residues 50-53, in the disulfide subloop of the dimer interface, but many other regions of the A4V protein exhibited hydrogen exchange properties identical to that of the wild type protein. Additionally, mass spectrometry revealed that A4V apoSOD1(S-S) undergoes slow localized unfolding in a large segment of the beta-barrel that included beta3, beta4, and loops II and III. In the disulfide-reduced form, A4V apoSOD1 exchanged like a "random coil" polypeptide at 20 degrees C and began to populate folded states at 4 degrees C. These local and global unfolding events could facilitate intermolecular protein-protein interactions that cause the aggregation or neurotoxicity of A4V SOD1.
authors.affiliation: Department of Chemistry and Biochemistry, University of California-Los Angeles, Los Angeles, CA 90095, USA.
authors.forename: Bryan Francis
authors.initials: BF
authors.lastname: Shaw
authors.forename: Armando
authors.initials: A
authors.lastname: Durazo
authors.forename: Aram M
authors.initials: AM
authors.lastname: Nersissian
authors.forename: Julian P
authors.initials: JP
authors.lastname: Whitelegge
authors.forename: Kym F
authors.initials: KF
authors.lastname: Faull
authors.forename: Joan Selverstone
authors.initials: JS
authors.lastname: Valentine
mesh_terms: D003080:Cold Temperature; D041961:Deuterium Exchange Measurement; D019281:Dimerization; D004220:Disulfides / Q000737:chemistry; D006801:Humans; D009154:Mutation; D017510:Protein Folding; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D021241:Spectrometry, Mass, Electrospray Ionization; D013482:Superoxide Dismutase / Q000737:chemistry* / Q000235:genetics*; D000072105:Superoxide Dismutase-1
chemical_list: D004220:Disulfides; C000606290:SOD1 protein, human; D013482:Superoxide Dismutase; D000072105:Superoxide Dismutase-1