title: Influence of protein disulfide isomerase (PDI) on antibody folding in vitro.
abstract: The role of eucaryotic protein disulfide isomerase (PDI) in the folding and reoxidation of proteins in vitro was investigated using an antibody Fab fragment as a model substrate, since PDI is known to participate in the disulfide bond formation of immunoglobulins in vivo. PDI has no effect on the folding of the Fab fragment with intact disulfide bonds, suggesting that, at least in this system, PDI is not able to influence the folding process in a chaperone-like manner. Instead, the role of PDI is limited to disulfide bond formation as demonstrated for the folding of the denatured and reduced Fab fragment. Here, PDI influences the yield of reactivation enormously with a maximum effect at about stoichiometric amounts of PDI and Fab. Furthermore, PDI changes the redox dependence of the reaction. In the presence of PDI, formation of the correct disulfide bonds is possible at higher oxidizing conditions compared to the spontaneous reaction. The requirements both for stoichiometric amounts of PDI and for the presence of PDI during the first seconds of refolding suggest that there is a kinetic competition between rapid structure formation of the antibody domains and interaction of PDI with cysteine residues in the folding protein.
authors.affiliation: Institut für Biophysik und Physikalische Biochemie, Universität Regensburg, Germany.
authors.forename: H
authors.initials: H
authors.lastname: Lilie
authors.forename: S
authors.initials: S
authors.lastname: McLaughlin
authors.forename: R
authors.initials: R
authors.lastname: Freedman
authors.forename: J
authors.initials: J
authors.lastname: Buchner
mesh_terms: D000818:Animals; D002417:Cattle; D004220:Disulfides; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000737:chemistry / Q000378:metabolism*; D007535:Isomerases / Q000378:metabolism*; D007700:Kinetics; D008099:Liver / Q000201:enzymology; D051379:Mice; D010084:Oxidation-Reduction; D019704:Protein Disulfide-Isomerases; D017510:Protein Folding
chemical_list: D004220:Disulfides; D007140:Immunoglobulin Fab Fragments; D007535:Isomerases; D019704:Protein Disulfide-Isomerases