Trypsinogen, refolding

Study of how the refolding of trypsinogen depends on the endogenous disulphide bridges... 

Fig. 5.18. Refolding of reduced Sepharose trypsinogen in different regeneration systems (from Sinha and Light, 1975) (A) 0.5 mM )5-mercaptoethanol (A) 0.5 mAf j -mercaptoe-thanol + 1 mM dehydroascorbic acid (O) 4 mM GSH + 0.4 mM GSSG. In all cases Tris 0.05 M CaCh buffer at pH 8.5, 35°C was used. The best regeneration mixture is the redox one containing 4 mM GSH and 0.4 mM GSSG.

Sinha and Light (1975) obtained better yield by using trypsinogen and trypsin immobilized on Agarose beads. With 0.2-0.6 mg of protein bound per ml of gel up to 60-70% regeneration yield was obtained in 24 h. Furthermore, the incorrectly folded structures continued to refold when placed in a mixture of reduced and oxidized glutathione to allow disulfide interchange. 

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