Site directed mutagenesis protein folding

In mutational T-value analyses of the transition state of protein folding, site-directed mutagenesis is used to introduce nondisruptive mutations at various... 

P204F <7> (<7> site-directed mutagenesis, mutation in the hinge region of the enzyme, which plays a role in protein folding during catalysis, less well folded with considerable loss of secondary and tertiary structure, no activity [59]) [59]... 

Site-directed mutagenesis can be used to establish the function of individual amino acid residues. This method also allows to introduce chemical probes (e.g., fluorophores) that may give insight into the dynamic features and folding properties of a flavoenzyme and its interaction with other protein partners (33, 34). In this context, it is important to stress that catalytically relevant conformational changes of flavoenzymes have been demonstrated that actually exploit the flavin itself as an excellent intrinsic spectroscopic probe (18, 31). 

Protein stability is the free energy difference (AG) between the folded and unfolded states at physiological conditions, and it is in the range of 5-25 kcal/mol. Site-directed mutagenesis experiments provided a wealth of data for understanding the importance of chemical interactions for the stability of proteins during amino acid substitutions. Protein stability is experimentally measured with differential scanning calorimetry, circular dichroism, fluorescence spectroscopy, and so forth. The availability of such data in an electronically accessible database would be a valuable resource for the analysis and prediction of protein mutant stability. 

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