THE REFOLDING OF HEN EGG WHITE RIBOFLAVIN-BINDING PROTEIN - EFFECT OF PROTEIN DISULPHIDE ISOMERASE ON THE REOXIDATION OF THE REDUCED PROTEIN

摘要

Hen egg white riboflavin-binding protein (RfBP) contains nine disulphide bonds. Provided these remain intact, the refolding of RfBP after incubation in 6 M guanidinium chloride is highly efficient with at least 95 % of the binding activity regained within 3 min. Kinetic studies indicate that this regain consists of at least two phases. When the disulphide bonds of RfBP are reduced, reoxidation using a mixture of oxidized and reduced glutathione leads to less than 5 % recovery of activity. However, if protein disulphide isomerase (PDI; EC 5.3.4.1) is present during the reoxidation nearly 50 % activity can be regained, suggesting that PDI may play an important role in the maturation of RfBP in vivo.