Evidence for a molten globule state as a general intermediate in protein folding

Abstract

The folding of globular proteins occurs through intermediate states whose characterisation provides information about the mechanism of folding. A major class of intermediate states is the compact ‘molten globule’, whose characteristics have been studied intensively in those conditions in which it is stable (at acid pH, high temperatures and intermediate concentrations of strong denaturants). In studies involving bovine carbonic anhydrase, human α-lact-albumin, bovine β-lactoglobulin, yeast phosphoglycerate kinase, β-lactamase from Staphylococcus aureus and recombinant human interleukin 1β, we have demonstrated that a transient intermediate which accumulates during refolding is compact and has the properties of the ‘molten globule’ state. We show that it is formed within 0.1–0.2 s. These proteins belong to different structural types (β, α + β and α/β), with and without disulphide bridges and they include proteins with quite different times of complete folding (from seconds to decades of minutes). We propose that the formation of the transient molten globule state occurs early on the pathway of folding of all globular proteins.