The aim of the proposed work is to study the folding of the B 1 domain of the immunoglobulin G binding protein from Streptococcus using a variety of experimental techniques, including stopped-flow, fluorescence, circular dicroism, urea, GuHCI and thermal denaturation, as well as site-directed mutagenesis. The B1 domain has the qualities to make it an ideal system for studying the folding process. Recently the E. coli strains super-expressing this domain have been designed by Dr. Serrano's group. This opens the possibility to start GB1 folding studies using all the techniques mentioned. In addition, we can introduce mutations into the domain surface to increase the stability of secondary structure elements. This influence can be a priori estimated by empirical algorithms developed by the group.
It would be most interesting to compare the observed experimental energetic changes with those predicted by theoretical calculations in order to investigate whether it is possible to use these algorithms for a rational increase in protein stability. The increase in protein stability is a major goal in protein engineering because of its potential industrial and pharmacological applications.