Investigating the in vivo folding kinetics and thermodynamics of phosphoglycerate kinase

The first chapter of this thesis introduces the field of protein folding. The occurrence of protein folding as a pathway instead of a random search is necessary for proteins to fold on biological time scales. It can be shown that small energetic biases towards a folded state can reduce the folding time of a protein to biologically relevant time scales. The study of protein folding seeks to understand the mechanism by which a protein transitions from an unfolded state to its native state. The recent development of many new experimental techniques including the fast relaxation imaging and temperature jumps used in our lab has allowed the study of protein folding in the cell. Chapter 2 of this thesis describes the development of a method to characterize variations in the folding of the protein phosphoglycerate kinase (PGK) inside living bacterial cells. Chapter 3 reports on the characterization of the spectral properties of CdSe quantum dots for use in shaping laser temperature jumps, so that kinetics of proteins in live cells can be studied with accurate temperature calibration.NSF MCB 1413256Ope