Chapter 13: Protein Folding, Maturation and Redox State

The direct protein expression approach described in Chapter 4 is ideally suited for observing cellular functional processes involving protein folding and maturation, cofactor binding and redox state regulation by in-cell NMR in human cells. In this chapter, some applications of in-cell NMR to investigate protein functional processes in HEK293T cells are described, including the maturation of superoxide dismutase 1, from zinc binding to copper delivery and disulphide bond formation, the effect of disease-linked mutations and the role of the specific copper chaperone, the redox-dependent folding of two small mitochondrial proteins as a function of specific redox partners and the effect of exogenous molecules on the protein redox state.