Effects of positively charged redox molecules on disulfide-coupled protein folding

• Published in: FEBS letters 2012-11, Vol.586 (21), p.3926-3930
• Main Authors: Okumura, Masaki, Shimamoto, Shigeru, Nakanishi, Takeyoshi, Yoshida, Yu-ichiro, Konogami, Tadafumi, Maeda, Shogo, Hidaka, Yuji
• Format: Article
• Language: English
• Subjects: 2-aminoethanethiol; 2-diethylaminoethanethiol; 2-dimethylaminoethanethiol; AET; arginyl-cysteinyl-glycine; ASA+; Chromatography, Reverse-Phase; DEAET; diisopropylaminoethanethiol; Disulfide; Disulfides - chemistry; DMAET; DPAET; Folding; Glutathione; Glutathione - chemistry; GSH and GSSG; Humans; Hydrogen-Ion Concentration; Intermediate; Kinetics; Lysozyme; MALDI-TOF/MS; matrix-assisted laser desorption ionization time of flight mass spectrometry; mercaptoethanol; mercaptopropionic acid; MPA; Muramidase - chemistry; Oxidation-Reduction; Protein Folding; Protein Precursors - chemistry; Prouroguanylin; RCG; reduced and oxidized form of glutathione, respectively; reversed-phase high performance liquid chromatography; RP-HPLC; Solutions; solvent accessible surface area with positive partial charges; Static Electricity; Sulfhydryl Compounds - chemistry; TFA; Thiol; trifluoroacetic acid; tris(hydroxymethyl)aminomethane hydrochloride; Tris/HCl
• ISSN: 0014-5793; 1873-3468
• DOI: 10.1016/j.febslet.2012.09.031

► Roles of cross-disulfide molecules in disulfide-coupled protein folding are proposed. ► Several types of aliphatic thiol compounds were examined for protein folding. ► Effects of the charge and molecular size of redox molecules were investigated. ► A larger ASA+ of thiol reagents is preferred for disulfide-coupled protein folding. In vitro folding of disulfide-containing proteins is generally regulated by redox molecules, such as glutathione. However, the role of the cross-disulfide-linked species formed between the redox molecule and the protein as a folding intermediate in the folding mechanism is poorly understood. In the present study, we investigated the effect of the charge on a redox molecule on disulfide-coupled protein folding. Several types of aliphatic thiol compounds including glutathione were examined for the folding of disulfide-containing-proteins, such as lysozyme and prouroguanylin. The results indicate that the positive charge and its dispersion play a critical role in accelerating disulfide-coupled protein folding.