Disulfide bond formation in the refolding of thermophilic fungal protein di
sulfide isomerase (PDI) was investigated. It was revealed that (i) a disulf
ide bond buried inside the molecule is preferentially formed and contribute
s to the thermal stability and the isomerizing power of PDI, and (ii) forma
tion of disulfide bonds in active sites located on the molecular surface ca
uses deformation of the optimum conformation resulting in a decrease in the
thermal stability.