title: Tuning Cooperativity on the Free Energy Landscape of Protein Folding.
abstract: Understanding the origin of the cooperativity seemingly inherent in a folding or unfolding reaction has been a major challenge. In particular, the relationship between folding cooperativity and stability is poorly understood. In this study, native state hydrogen exchange in conjunction with mass spectrometry has been used to explore the free energy landscape accessible to the small protein monellin, when the stability of the protein is varied. Mass distributions obtained in the EX1 limit of exchange have allowed a direct distinction between correlated (cooperative) and uncorrelated (noncooperative) structure-opening processes. Under conditions where the native protein is maximally stable, a continuum of partially unfolded states is gradually sampled before the globally unfolded state is transiently sampled. Under conditions that stabilize the unfolded state of the protein, the slowest structure-opening reactions leading to complete unfolding become cooperative. The present study provides experimental evidence for a gradual uphill unfolding transition on a very slow time scale, in the presence of a large free energy difference between the native and unfolded states. The results suggest that the cooperativity that manifests itself in protein folding and unfolding reactions carried out in the presence of denaturant might merely be a consequence of the effect of the denaturant on the unfolded state and transition state stabilities.
authors.affiliation: National Centre for Biological Sciences, Tata Institute of Fundamental Research, Bengaluru 560065, India.
authors.forename: Pooja
authors.initials: P
authors.lastname: Malhotra
authors.affiliation: National Centre for Biological Sciences, Tata Institute of Fundamental Research, Bengaluru 560065, India.
authors.forename: Jayant B
authors.initials: JB
authors.lastname: Udgaonkar
mesh_terms: D008956:Models, Chemical; D010940:Plant Proteins / Q000737:chemistry*; D017510:Protein Folding; D017434:Protein Structure, Tertiary
chemical_list: D010940:Plant Proteins; C002541:monellin protein, Dioscoreophyllum cumminsii