Different folding transition states may result in the same native structure.

The crystal structures of two circular permutants of the alpha-spectrin SH3 domain with new termini within the RT loop (S19-P20s) and the distal loop (N47-D48s) have been determined at 2.02 and 1.77 A resolution respectively. Both fold into the same three-dimensional structure as the wild-type SH3 domain except for the engineered loop that fuses the wild-type termini. The cleaved RT loop in S19-P20s loses nine conserved hydrogen bonds through local hydrogen bond unzipping; no hydrogen bond unzipping occurs in N47-D48s. The structures of the transition states for folding of wild-type alpha-spectrin SH3 domain and the two circular permutants have been examined by analysis of the folding kinetics of eight strategically distributed point mutants. Unlike the native structures, the transition states of the three proteins are considerably different, suggesting that there is no direct relationship between these two states in a protein.