Effects of conserved alanine to glycine substitutions on equilibrium stability. CD-monitored urea-induced unfolding transitions of constructs that contain alanine to glycine subsitutions in Nank1-5 (A), Nank1-5C1 (B), and Nank1-5C2 (C). Fluorescence-monitored transitions of substitutions in Nank1-5C2 (D). The position of each substitution is indicated as follows: first repeat, red; second repeat, orange; third repeat, yellow; fourth repeat, green; fifth repeat, blue; first designed, purple; second designed repeat, light purple.