NMR Assignments of the b′ and a′ Domains of Thermophilic Fungal Protein Disulfide Isomerase

Redox-Dependent Domain Rearrangement of Protein Disulfide Isomerase Coupled with Exposure of Its Substrate-Binding Hydrophobic Surface

“…2) on the basis of the NMR spectral assignments reported previously. 23 Although the solution structures of these domains are similar to those of their counterparts in the crystal structure of yeast PDI (r.m.s.d. of 1.8 Å for b′ domain and 1.9 Å for a′ domain), the b′ domain of thermophilic fungal PDI lacks the short helical segment (α5) preceding the last helix observed for yeast PDI (Supplementary Fig.…”

“…23 Full-length PDI (1-485) and a Cterminally truncated mutant a-b fragment (1-219) were expressed with an N-terminal hexahistidine tag and purified by Ni 2+ -NTA affinity chromatography, while an N-terminally truncated mutant b′-a′ fragment (208-449) was produced with an N-terminal glutathione Stransferase tag and purified by glutathione-Sepharose chromatography. After removal of the tags, the proteins were further purified as described previously.…”

“…After removal of the tags, the proteins were further purified as described previously. 23 Oxidized PDI and b′-a′ oxi were prepared by dialyzing the sample against 50 mM Tris-HCl (pH 8.0), including 0.1 mM oxidized glutathione, at 277 K for a week.…”

Redox-dependent conformational transition of catalytic domain of protein disulfide isomerase indicated by crystal structure-based molecular dynamics simulation

“…The purification of reduced and oxidized b′-a′ domains (residues 208−449) from Humicola insolens was performed as previously described [8,15]. For crystallization, the reduced and oxidized proteins, dissolved in 10 mM Tris-HCl (pH 8.0) containing 10 mM dithiothreitol (DTT) or 0.1 mM oxidized glutathione, respectively, were concentrated to 15 mg/mL.…”

Protein Disulfide Isomerase: Structure, Mechanism of Oxidative Protein Folding and Multiple Functional Roles

“…The structural and functional studies on PDI from various species, including human [12][13][14][15][16] yeast [17,18] , and thermophilic fungus [19][20][21] have been reported by many researchers. The data showed that PDI consists of four tandem thioredoxin-like domains a, b, b′, and a′ plus a C-terminal extension [3][4][5]22] , which are arranged into a U-shape structure [16][17][18] .…”