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Protein folding research often finds complex kinetic behavior which shows that unfolded molecules take several stages to convert to the fully native form. A Pro- variate of SNase's unfolding and folding kinetic mechanism was characterized using stopped-flow fluorescence methods, with alanines and glycines used in place of six proline residues. The folding kinetics still demonstrate a complex denaturant dependence, even though complicating factors such as prolines have been removed. A sequential four-state folding mechanism can be used to model the primary stages in unfolding and folding.

The backbone 1H and 15N resonances of unligated staphylococcal nuclease H124L were assigned by three-dimensional (3D) 1H-15N nuclear Overhauser- multiple-quantum coherence (NOESY-HMQC) nuclear magnetic resonance spectroscopy. Secondary structural elements (alpha-helices, beta-sheets, reverse turns and loops) were determined by analysis of patterns of NOE connectivities present in the 3D spectrum.