title: Chemical methods for producing disulfide bonds in peptides and proteins to study folding regulation.
abstract: Disulfide bonds play a critical role in the folding of secretory and membrane proteins. Oxidative folding reactions of disulfide bond-containing proteins typically require several hours or days, and numerous misbridged disulfide isomers are often observed as intermediates. The rate-determining step in refolding is thought to be the disulfide-exchange reaction from nonnative to native disulfide bonds in folding intermediates, which often precipitate during the refolding process because of their hydrophobic properties. To overcome this, chemical additives or a disulfide catalyst, protein disulfide isomerase (PDI), are generally used in refolding experiments to regulate disulfide-coupled peptide and protein folding. This unit describes such methods in the context of the thermodynamic and kinetic control of peptide and protein folding, including (1) regulation of disulfide-coupled peptides and protein folding assisted by chemical additives, (2) reductive unfolding of disulfide-containing peptides and proteins, and (3) regulation of disulfide-coupled peptide and protein folding using PDI.
authors.affiliation: Institute of Multidisciplinary Research for Advanced Materials, Tohoku University, Miyagi, Japan.
authors.forename: Masaki
authors.initials: M
authors.lastname: Okumura
authors.affiliation: Faculty of Science and Engineering, Kinki University, Osaka, Japan.
authors.forename: Shigeru
authors.initials: S
authors.lastname: Shimamoto
authors.affiliation: Faculty of Science and Engineering, Kinki University, Osaka, Japan.
authors.forename: Yuji
authors.initials: Y
authors.lastname: Hidaka
mesh_terms: D004220:Disulfides / Q000737:chemistry*; D010455:Peptides / Q000737:chemistry*; D019704:Protein Disulfide-Isomerases / Q000737:chemistry*; D058767:Protein Unfolding; D011506:Proteins / Q000737:chemistry*
chemical_list: D004220:Disulfides; D010455:Peptides; D011506:Proteins; D019704:Protein Disulfide-Isomerases
keywords: additive; disulfide; folding; glutathione; protein disulfide isomerase