NATURE AND CONSEQUENCES OF GROEL-PROTEIN INTERACTIONS

The importance of chaperonin-protein interactions has been investigate d by analyzing the refolding of the barley chymotrypsin inhibitor 2 in the presence of GroEL. The chaperonin retards the rate of refolding o f wild type and 32 representative point mutants. The retardation of th e rate drops to a finite level at saturating concentrations of GroEL, being lowered by a factor of 3-100, depending on the mutation. It is s een qualitatively that truncation of large hydrophobic side chains to smaller side chains weakens binding. Analysis of the magnitude of the rates of retardation shows further that hydrophobic and positively cha rged side chains tend to interact favorably with GroEL whereas negativ ely charged side chains tend to repel. There is an inverse correlation between the strength of hydrophobic interactions and the rate constan t for refolding of the GroEL-complexed protein: the better the binding , the slower the folding. This shows directly that hydrophobic (and ot her favorable) interactions between the chaperonin and substrate are w eakened during the refolding process and implies that unfolding can be catalyzed by the gain of such interactions.