title: Cooperation of ER-60 and BiP in the oxidative refolding of denatured proteins in vitro.
abstract: ER-60 is a PDI family protein that has protein thiol-disulfide oxidoreductase activity. It has been shown to associate with BiP in the endoplasmic reticulum. Here, we analyzed the cooperation of ER-60 and BiP in the oxidative refolding of denatured proteins in vitro. ER-60 facilitated the refolding of 20 or 30% of denatured alpha-lactalbumin or ribonuclease B during incubation for 80 min, and these levels of nearly doubled on the addition of BiP to the reaction mixture. BiP alone could not refold denatured ribonuclease B, but could refold denatured alpha-lactalbumin a little. Enhancement of oxidative refolding of alpha-lactalbumin by ER-60 could be detected only when ER-60 was present from the start of refolding. On surface plasmon resonance analysis, ER-60 was shown to associate with BiP. The association was not influenced by ATP or ADP. Domains a and/or b' of ER-60 were implicated in the association with BiP.
authors.affiliation: Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Gokasho, Uji, Kyoto 611-0011.
authors.forename: Hirokazu
authors.initials: H
authors.lastname: Okudo
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Kato
authors.forename: Yukino
authors.initials: Y
authors.lastname: Arakaki
authors.forename: Reiko
authors.initials: R
authors.lastname: Urade
mesh_terms: D000091342:Endoplasmic Reticulum Chaperone BiP; D006360:Heat-Shock Proteins / Q000737:chemistry*; D006801:Humans; D007768:Lactalbumin; D018832:Molecular Chaperones / Q000737:chemistry*; D011485:Protein Binding; D019704:Protein Disulfide-Isomerases / Q000737:chemistry*; D017510:Protein Folding; D020673:Protein Renaturation; D012260:Ribonucleases; D020349:Surface Plasmon Resonance
chemical_list: D000091342:Endoplasmic Reticulum Chaperone BiP; D006360:Heat-Shock Proteins; D018832:Molecular Chaperones; D007768:Lactalbumin; D012260:Ribonucleases; C029600:ribonuclease B; D019704:Protein Disulfide-Isomerases; C085868:PDIA3 protein, human