title: Temporal development of protein structure during S100A11 folding and dimerization probed by oxidative labeling and mass spectrometry.
abstract: Considerable progress in deciphering the mechanisms of protein folding has been made. However, most work in this area has focused on single-chain systems, whereas the majority of proteins are oligomers. The spontaneous assembly of intact multi-subunit systems from disordered building blocks encompasses the formation of intramolecular as well as intermolecular contacts. Both types of interaction affect the solvent accessibility of individual protein segments. This work employs pulsed hydroxyl radical (·OH) labeling for tracking time-dependent accessibility changes during folding and assembly of the S100A11 homodimer. ·OH induces covalent modifications at exposed residues. Structural snapshots are obtained by combining ·OH labeling with rapid mixing and mass spectrometry. The free subunits are found to possess a partially non-native hydrophobic core that prevents subunit association during the initial stages of the reaction. Instead, the protein forms an early (10 ms) monomeric intermediate that exhibits reduced solvent accessibility in regions distant from helices I and IV, which constitute the dimerization interface. Subunit association is complete after 800 ms, although the protein retains significant disorder in helices II and III at this point. Subsequent consolidation of these elements leads to the native state. The experimental strategy used here could become a general tool for deciphering kinetic mechanisms of biomolecular self-assembly processes.
authors.affiliation: Department of Biochemistry, The University of Western Ontario, London, Ontario, Canada N6A 5C1.
authors.forename: Bradley B
authors.initials: BB
authors.lastname: Stocks
authors.forename: Atoosa
authors.initials: A
authors.lastname: Rezvanpour
authors.forename: Gary S
authors.initials: GS
authors.lastname: Shaw
authors.forename: Lars
authors.initials: L
authors.lastname: Konermann
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D019281:Dimerization; D013058:Mass Spectrometry / Q000379:methods*; D008958:Models, Molecular; D010084:Oxidation-Reduction; D011487:Protein Conformation; D017510:Protein Folding; D011817:Rabbits; D009418:S100 Proteins / Q000737:chemistry* / Q000235:genetics
chemical_list: D009418:S100 Proteins