The folding catalyst protein disulfide isomerase is constructed of active and inactive thioredoxin modules

AbstractBackground: Protein disulfide isomerase (PDI), a multifunctional protein of the endoplasmic reticulum, catalyzes the formation, breakage and rearrangement of disulfide bonds during protein folding. Dissection of this protein into its individual domains has confirmed the presence of the a and a′ domains, which are homologous to thioredoxin, having related structures and activities. The a and a′ domains both contain a –Cys–Gly–His–Cys– active-site sequence motif. The remainder of the molecule consists primarily of two further domains, designated b and b′ which are thought to be sequence repeats on the basis of a limited sequence similarity. The functions of the b and b′ domains are unknown and, until now, the structure of neither domain was known.Results: Heteronuclear nuclear magnetic resonance (NMR) methods have been used to determine the global fold of the PDI b domain. The protein has an α/β fold with the order of the elements of secondary structure being β1–α1–β2–α2–β3–α3–β4–β5–α4. The strands are all in a parallel arrangement with respect to each other, except for β4 which is antiparallel. The arrangement of the secondary structure elements of the b domain is identical to that found in the a domain of PDI and in the ubiquitous redox protein thioredoxin; the three-dimensional folding topology of the b domain is also very similar to that of these proteins.Conclusions: Our determination of the global fold of the b domain of PDI by NMR reveals that, like the a domain, the b domain contains the thioredoxin motif, even though the b domain has no significant amino-acid sequence similarities to any members of the thioredoxin family. This observation, together with indications that the b′ domain adopts a similar fold, suggests that PDI consists of active and inactive thioredoxin modules. These modules may have been adapted during evolution to provide PDI with its complete spectrum of enzymatic activities